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Stronach, B. E., S. E. Siegrist i M. C. Beckerle. "Two muscle-specific LIM proteins in Drosophila." Journal of Cell Biology 134, nr 5 (1.09.1996): 1179–95. http://dx.doi.org/10.1083/jcb.134.5.1179.
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The LIM domain defines a zinc-binding motif found in a growing number of eukaryotic proteins that regulate cell growth and differentiation during development. Members of the cysteine-rich protein (CRP) family of LIM proteins have been implicated in muscle differentiation in vertebrates. Here we report the identification and characterization of cDNA clones encoding two members of the CRP family in Drosophila, referred to as muscle LIM proteins (Mlp). Mlp60A encodes a protein with a single LIM domain linked to a glycine-rich region. Mlp84B encodes a protein with five tandem LIM-glycine modules. In the embryo, Mlp gene expression is spatially restricted to somatic, visceral, and pharyngeal muscles. Within the somatic musculature, Mlp84B transcripts are enriched at the terminal ends of muscle fibers, whereas Mlp60A transcripts are found throughout the muscle fibers. The distributions of the Mlp60A and Mlp84B proteins mirror their respective mRNA localizations, with Mlp84B enrichment occurring at sites of muscle attachment. Northern blot analysis revealed that Mlp gene expression is developmentally regulated, showing a biphasic pattern over the course of the Drosophila life cycle. Peaks of expression occur late in embryogenesis and during metamorphosis, when the musculature is differentiating. Drosophila Mlp60A and Mlp84B, like vertebrate members of the CRP family, have the ability to associate with the actin cytoskeleton when expressed in rat fibroblast cells. The temporal expression and spatial distribution of muscle LIM proteins in Drosophila are consistent with a role for Mlps in myogenesis, late in the differentiation pathway.
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Wishard, Rohan, Mohan Jayaram, Saraf R. Ramesh i Upendra Nongthomba. "Spatial and temporal requirement of Mlp60A isoforms during muscle development and function in Drosophila melanogaster". Experimental Cell Research 422, nr 1 (styczeń 2023): 113430. http://dx.doi.org/10.1016/j.yexcr.2022.113430.
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Nadia, Zubyda Mushtari, Prosun Roy, Sayed Mashequl Bari i Md Abdus Salam. "Dietary effect of moringa (Moringa oleifera; Lamarck, 1785) leaf powder on growth response of tilapia (Oreochromis niloticus; Linnaeus, 1758)". Asian Journal of Medical and Biological Research 7, nr 2 (30.06.2021): 153–63. http://dx.doi.org/10.3329/ajmbr.v7i2.54995.
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Management of fish health is one of the main considerations in aquaculture and different plant compounds are being used for supporting fish health to minimize the negative impacts of synthetic aqua drugs. In the present experiment, potentiality of moringa (Moringa oleifera; Lamarck, 1785) leaf as a nutritious dietary source for tilapia (Oreochromis niloticus; Linnaeus, 1758) fish was tested and the duration was two months from 30th September to 30th November, 2020. The moringa leaves were brought, cleaned, dried, and finally crushed into powder. Three experimental diets were formulated using the processed moringa leaf powder (MLP) at the rate of 0% (MLP0%) as control, 10% (MLP10%) and 20% (MLP20%) as treatment mixing with rice bran, wheat bran, mustard oil cake, fish meal, soya oil and vitamin-mineral premix. Fifteen tilapia fingerlings having average initial length 10.88±0.11 cm and initial weight 29.06±0.50 g was stocked in each tank with 90 L water. Sixty days feeding trial was performed with three replications of each treatment. The fishes were fed with formulated feeds twice daily at 9 am and 4 pm at a rate of 3% of their body mass. Sampling of fish and water quality parameters were carried out at twelve days interval. Moreover, the blood glucose and cholesterol of tilapia were measured monthly. Final length, final weight, weight gain, percent weight gain, feed conversion ratio (FCR), specific growth rate (SGR) and production of tilapia were significantly different among the treatments. The highest FCR (3.17±0.25) and SGR (1.33±0.12 %) values were in MLP20% and MLP10%, respectively. In the experiment, the highest and the lowest tilapia production were 9.21±0.39 and 7.39±0.35 kg m-3 in MLP10% and MLP20%, respectively. The blood glucose values were significantly different among the treatments (p< 0.05) and the highest value was in MLP0% (48.00±2.00 mg dl-1). Moreover, the highest and the lowest blood cholesterol was found in MLP0% (177.67±2.52 mg dl-1) and MLP20% (148.33±1.53 mg dl-1), respectively whereas the values were highly significantly different among the treatments (p≤ 0.01). Water quality parameters were statistically similar among the treatments (p> 0.05) and the values were within acceptable range for tilapia culture. Asian J. Med. Biol. Res. 2021, 7 (2), 153-163
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Stronach, Beth E., Patricia J. Renfranz, Brenda Lilly i Mary C. Beckerle. "Muscle LIM Proteins Are Associated with Muscle Sarcomeres and Require dMEF2 for Their Expression during DrosophilaMyogenesis". Molecular Biology of the Cell 10, nr 7 (lipiec 1999): 2329–42. http://dx.doi.org/10.1091/mbc.10.7.2329.
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A genetic hierarchy of interactions, involving myogenic regulatory factors of the MyoD and myocyte enhancer-binding 2 (MEF2) families, serves to elaborate and maintain the differentiated muscle phenotype through transcriptional regulation of muscle-specific target genes. Much work suggests that members of the cysteine-rich protein (CRP) family of LIM domain proteins also play a role in muscle differentiation; however, the specific functions of CRPs in this process remain undefined. Previously, we characterized two members of the Drosophila CRP family, the muscle LIM proteins Mlp60A and Mlp84B, which show restricted expression in differentiating muscle lineages. To extend our analysis ofDrosophila Mlps, we characterized the expression of Mlps in mutant backgrounds that disrupt specific aspects of muscle development. We show a genetic requirement for the transcription factor dMEF2 in regulating Mlp expression and an ability of dMEF2 to bind, in vitro, to consensus MEF2 sites derived from those present inMlp genomic sequences. These data suggest that theMlp genes may be direct targets of dMEF2 within the genetic hierarchy controlling muscle differentiation. Mutations that disrupt myoblast fusion fail to affect Mlp expression. In later stages of myogenic differentiation, which are dedicated primarily to assembly of the contractile apparatus, we analyzed the subcellular distribution of Mlp84B in detail. Immunofluorescent studies revealed the localization of Mlp84B to muscle attachment sites and the periphery of Z-bands of striated muscle. Analysis of mutations that affect expression of integrins and α-actinin, key components of these structures, also failed to perturb Mlp84B distribution. In conclusion, we have used molecular epistasis analysis to position Mlp function downstream of events involving mesoderm specification and patterning and concomitant with terminal muscle differentiation. Furthermore, our results are consistent with a structural role for Mlps as components of muscle cytoarchitecture.
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Amaro, V., S. Cavuoti, M. Brescia, C. Vellucci, C. Tortora i G. Longo. "METAPHOR: Probability density estimation for machine learning based photometric redshifts". Proceedings of the International Astronomical Union 12, S325 (październik 2016): 197–200. http://dx.doi.org/10.1017/s1743921317002186.
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AbstractWe present METAPHOR (Machine-learning Estimation Tool for Accurate PHOtometric Redshifts), a method able to provide a reliable PDF for photometric galaxy redshifts estimated through empirical techniques. METAPHOR is a modular workflow, mainly based on the MLPQNA neural network as internal engine to derive photometric galaxy redshifts, but giving the possibility to easily replace MLPQNA with any other method to predict photo-z’s and their PDF. We present here the results about a validation test of the workflow on the galaxies from SDSS-DR9, showing also the universality of the method by replacing MLPQNA with KNN and Random Forest models. The validation test include also a comparison with the PDF’s derived from a traditional SED template fitting method (Le Phare).
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Hao Yuan, Hao Yuan, Xiao Meng Hao Yuan, Kai Xu Xiao Meng i Qing Jia Kai Xu. "A Practical Machine-Learning-Based Approach for Leather Automatic Defect Inspection". 電腦學刊 33, nr 5 (październik 2022): 019–28. http://dx.doi.org/10.53106/199115992022103305002.
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<p>Leather manual inspection is common in many industries, these methods are low efficiency and cannot be in line with automated manufacturing. In this paper, we propose a leather automated defect inspection (LADI) method based on machine learning and establish a practical LADI system composed of four modules: image acquisition, image preprocessing, image segmentation, and post-processing. The LADI method which forms the image segmentation module is a combination of multi-layer perceptron (MLP) and principal component analysis (PCA), namely MLPPCA. We propose two new algorithms that image preprocessing and post-processing to enhance the image quality and enrich details of the segmentation result. In the result analysis, compare MLPPCA, MLP, KNN, SVMRBF, GMM, show that MLPPCA has strong competitiveness in performance and execution time. The LADI system has been used in a China leather factory, the feedback shows that it combines the advantages of high inspection accuracy and short execution time.</p> <p> </p>
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Cifuentes, Rosa, José Padilla, María Eugenia de la Morena-Barrio, Belén de la Morena-Barrio, Carlos Bravo-Pérez, Pedro Garrido-Rodríguez, María Llamas i in. "Usefulness and Limitations of Multiple Ligation-Dependent Probe Amplification in Antithrombin Deficiency". International Journal of Molecular Sciences 24, nr 5 (6.03.2023): 5023. http://dx.doi.org/10.3390/ijms24055023.
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Multiplex ligation-dependent probe amplification (MLPA) identifies genetic structural variants in SERPINC1 in 5% of cases with antithrombin deficiency (ATD), the most severe congenital thrombophilia. Our aim was to unravel the utility and limitations of MLPA in a large cohort of unrelated patients with ATD (N = 341). MLPA identified 22 structural variants (SVs) causing ATD (6.5%). MLPA did not detect SVs affecting introns (four cases), and the diagnosis was inaccurate in two cases according to long-range PCR or nanopore sequencing. MLPA was used to detect possible hidden SVs in 61 cases with type I deficiency with single nucleotide variations (SNVs) or small insertion/deletion (INDEL). One case had a false deletion of exon 7, as the 29-bp deletion affected an MLPA probe. We evaluated 32 variants affecting MLPA probes: 27 SNVs and 5 small INDELs. In three cases, MLPA gave false-positive results, all diagnosed as deletions of the affected exon: a small INDEL complex, and two SNVs affecting MLPA probes. Our study confirms the utility of MLPA to detect SVs in ATD, but also shows some limitations in detecting intronic SVs. MLPA renders imprecise and false-positive results for genetic defects which affect MLPA probes. Our results encourage the validation of MLPA results.
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Guo, Leiming, Kun Shi i Lin Wang. "MLPMDA: Multi-layer linear projection for predicting miRNA-disease association". Knowledge-Based Systems 214 (luty 2021): 106718. http://dx.doi.org/10.1016/j.knosys.2020.106718.
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Alzahrani, Ali, i Md Al-Amin Bhuiyan. "Feature selection for urban land cover classification employing genetic algorithm". Bulletin of Electrical Engineering and Informatics 11, nr 2 (1.04.2022): 793–802. http://dx.doi.org/10.11591/eei.v11i2.3399.
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Feature selection has attained substantial research interest in image processing, computer vision, pattern recognition and so on due to tremendous dimensional reduction in image analysis. This research addresses a genetic algorithm based feature selection strategy for urban land cover classification. The principal purpose of this research is to monitor the land cover alterations in satellite imagery for urban planning. The method is based on object based classification by detecting the object area of a given image with the knowledge of visual information of the object from remote sensing images. The classification system is organized through a multilayer perceptron with genetic algorithm (MLPGA). Experimental results explicitly indicate that this MLPGA based hybrid feature selection procedure performs classification with sensitivity 94%, specificity 90% and precision 89%, respectively. This MLPGA centered hybrid feature selection scheme attains better performance than the counterpart methods in terms of classification accuracy.
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Lin, Lin, Changsheng Tong, Feng Guo, Song Fu, Yancheng Lv i Wenhui He. "A Self-Attention Integrated Learning Model for Landing Gear Performance Prediction". Sensors 23, nr 13 (7.07.2023): 6219. http://dx.doi.org/10.3390/s23136219.
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The landing gear structure suffers from large loads during aircraft takeoff and landing, and an accurate prediction of landing gear performance is beneficial to ensure flight safety. Nevertheless, the landing gear performance prediction method based on machine learning has a strong reliance on the dataset, in which the feature dimension and data distribution will have a great impact on the prediction accuracy. To address these issues, a novel MCA-MLPSA is developed. First, an MCA (multiple correlation analysis) method is proposed to select key features. Second, a heterogeneous multilearner integration framework is proposed, which makes use of different base learners. Third, an MLPSA (multilayer perceptron with self-attention) model is proposed to adaptively capture the data distribution and adjust the weights of each base learner. Finally, the excellent prediction performance of the proposed MCA-MLPSA is validated by a series of experiments on the landing gear data.
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Crauciuc, George Andrei, Florin Tripon, Alina Bogliş, Amalia Făgărăşan i Claudia Bănescu. "Multiplex ligation dependent probe amplification - A useful, fast and cost-effective method for identification of small supernumerary marker chromosome in children with developmental delay and congenital heart defect". Revista Romana de Medicina de Laborator 26, nr 4 (1.10.2018): 461–70. http://dx.doi.org/10.2478/rrlm-2018-0032.
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Abstract Small supernumerary marker chromosome (sSMC) is a rare chromosomal abnormality and is detected in about 0.3% in cases with multiple congenital anomalies (MCA) and/or developmental delay. Different techniques for investigation of cases with MCA and/or developmental delay are available ranging from karyotyping to molecular cytogenetic technique and ultimately multiplex ligation dependent probe amplification (MLPA). Here we present a patient with multiple congenital anomalies for which classical cytogenetic technique was used as a first step in diagnosis and the results being confirmed by MLPA. The karyotype disclosed a sSMC considered to be a fragment of chromosome 22. The MLPA analysis using SALSA MLPA probemix P064-C2 Microdeletion Syndromes-1B confirmed the karyotype results, and according to the manufacturer’s recommendation we performed another confirmation analysis with MLPA probemix P311-B1 Congenital Heart Disease and MLPA probemix P250-B2 DiGeorge. We also suspected an Emanuel syndrome and performed another MLPA analysis with SALSA MLPA probemix P036-E3 Subtelomeres Mix 1 and probemix P070-B3 Subtelomeres Mix 2B for investigation of subtelomeric region that revealed a duplication of 11q25 region and the confirmation was performed using SALSA MLPA probemix P286-B2 Human Telomere-11. In conclusion, we consider that MLPA is a valuable method for identification of sSMC in children with developmental delay and congenital anomalies. Genetic diagnosis using different molecular techniques, such as MLPA, for increasing accuracy in identification of chromosomal structural aberrations has an important role in clinical diagnosis and in genetic counselling and our case explain the importance of using a specific laboratory technique for each stage of diagnosis.
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Marcinkowska, Malgorzata, Kwok-Kin Wong, David J. Kwiatkowski i Piotr Kozlowski. "Design and Generation of MLPA Probe Sets for Combined Copy Number and Small-Mutation Analysis of Human Genes: EGFR as an Example". Scientific World JOURNAL 10 (2010): 2003–18. http://dx.doi.org/10.1100/tsw.2010.195.
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Multiplex ligation-dependent probe amplification (MLPA) is a multiplex copy number analysis method that is routinely used to identify large mutations in many clinical and research labs. One of the most important drawbacks of the standard MLPA setup is a complicated, and therefore expensive, procedure of generating long MLPA probes. This drawback substantially limits the applicability of MLPA to those genomic regions for which ready-to-use commercial kits are available. Here we present a simple protocol for designing MLPA probe sets that are composed entirely of short oligonucleotide half-probes generated through chemical synthesis. As an example, we present the design and generation of an MLPA assay for parallel copy number and small-mutation analysis of theEGFRgene.
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DAMATO, BE, J. DOPIERALA i SE COUPLAND. "MLPA of choroidal melanoma". Acta Ophthalmologica 88 (wrzesień 2010): 0. http://dx.doi.org/10.1111/j.1755-3768.2010.4262.x.
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COUPLAND, S., M. VAN DIJK, J. SIBBRING, P. HOWARD i B. DAMATO. "MLPA of uveal melanomas". Acta Ophthalmologica Scandinavica 85 (2.10.2007): 0. http://dx.doi.org/10.1111/j.1600-0420.2007.01063_3552.x.
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COUPLAND, S., i B. DAMATO. "MLPA of extraocular tumors". Acta Ophthalmologica Scandinavica 85 (2.10.2007): 0. http://dx.doi.org/10.1111/j.1600-0420.2007.01063_3553.x.
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Velloso, Elvira D. Rodrigues Pereira, Camila L. Clozato, Mauren Fernanda Muller Santos, Maria Gabriella Cordeiro, Roberta Sitnik, Joao Renato R. Pinho i Cristovao Luis P. Mangueira. "Complementary Molecular Biology Techniques for the Detection of Unbalanced Chromosomal Abnormalities in Acute Myeloid Leukemias and Myelodysplastic Syndromes: Comparison of Fluorescence In Situ Hybridization (FISH) and Multiplex Ligation-Dependent Probe Amplification (MLPA)". Blood 128, nr 22 (2.12.2016): 5525. http://dx.doi.org/10.1182/blood.v128.22.5525.5525.
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Abstract Background: The accurate detection of cytogenetic abnormalities in Acute Myeloid Leukemias (AML) and Myelodysplastic Syndromes (MDS) play an important role in prognostic value and often assist in therapeutic choice. The most commonly used genetic test for the diagnostics of these conditions is karyotype (KT), which is able to detect frequent non-random chromosomal abnormalities. However, this technique requires metaphases and offers low resolution (5Mb), which may lead to the loss of clinically important smaller genomic rearrangements. Therefore, a complementary technique is sometimes needed. FISH, for example, is a highly reliable technique, it can be performed in interphases, has higher resolution, and can detect balanced translocations (important for prognosis mainly in AML), although it is very expensive. As an alternative method, MLPA is usually less costly, is able to evaluate more genomic regions at once and is performed with genomic DNA, which is convenient for biological material availability. On the other hand, it is not able to detect balanced translocations due to its semi-quantitative PCR-based method. The aim of this study was to evaluate MLPA as a complementary technique to KT, and to compare the outcomes of MLPA and FISH panels for the high resolution detection of common unbalanced genetic alterations in MDS and AML. Methods: A total of 23 samples from patients diagnosed with MDS (n=16)/AML(n=6) were tested for KT and MLPA. 20 metaphases were analyzed in G-banding KT. MLPA (Kits P144 and P145, MRC-Holland) was performed with genomic DNA extracted from bone marrows. A subset of 10 samples was tested for FISH MDS/AML panel (PML/RARA, CBFB/MYH11, RUNX1/RUNX1T1, MLL rearrangement, del5q, del7q, de17p and del20q, Cytocell), and the outcomes were compared to MLPA. Reagent costs were also compared between MLPA and FISH techniques. Results: Outcomes from KT and MLPA were mostly concordant: 91.3% of samples had either concordant or partially concordant results (considering "partially concordant" when MLPA could detect smaller genetic alterations not achieved by KT). Discordance between KT and MLPA occurred in two cases: (i) MLPA detected a deletion of RUNX1 gene (chr 21) in a previously normal KT, and (ii) MLPA showed different abnormalities than those in a complex KT case, with low mitotic index. When comparing regions that are notoriously related to MDS/AML (-5/del5q, -7/del7q, +8, del11q, del13q, del17p, del20q and del21q), all results were concordant between MLPA and KT. Comparison of MLPA and FISH was performed with 10 samples, of which 4 were negative for genetic alterations and 6 were positive. 90% of samples had concordant results, and 10% (one sample) showed extra genetic alterations in MLPA (del7q; del11q and del20q). This is expected since MLPA analyzes more genetic regions then FISH MDS/AML panel at once. Genomic regions accessed both by MLPA and FISH included -7, +8, del17p, del21q. Regarding financial expenses (cost of commercial MDS/AML panels and additional reagents), MLPA costs approximately U$280.00 (considering test sample plus internal controls), while FISH costs U$555.00 (values were converted from quotes provided by regional distributors and converted to dollars). Conclusions: MLPA results are equivalent to FISH for the detection of MDS/AML frequent unbalanced genetic alterations. Both techniques are reliable as complementary molecular biology tests for KT. Moreover, MLPA is a friendly and less expensive technique for the detection of high resolution genomic unbalanced abnormalities, frequently adding extra information to KT results. Nevertheless, it is important to notice that only FISH can detect balanced translocations with significant prognostic value in AML. Disclosures No relevant conflicts of interest to declare.
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Гималова, Г. Ф., З. С. Абдуллин i Э. К. Хуснутдинова. "MLPA-analysis of TP53 gene in patients with small-cell lung cancer". Nauchno-prakticheskii zhurnal «Medicinskaia genetika», nr 6(215) (29.06.2020): 83–85. http://dx.doi.org/10.25557/2073-7998.2020.06.83-85.
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Мелкоклеточный рак легкого (МРЛ) составляет около 15-20% всех случаев РЛ и относится к наиболее злокачественно текущим опухолям. В более чем 90 % случаев МРЛ выявляются мутации гена TP53. Целью нашего исследования являлся анализ структурных изменений хромосомной области 17p13 у больных МРЛ из Республики Башкортостан. Материалом исследования служили 70 образцов тканей легких больных МРЛ. Методом исследования являлся MLPA-анализ. Делеции и/или дупликации в кодирующих областях гена TP53 выявлены в 40% образцов, в т.ч. дупликации первого (23% образцов) и делеции 4-6-го и 11-го экзонов гена TP53 (17% образцов). Кроме того, в четырех образцах выявлена мутация c.1100delC гена CHEK2. Small cell lung cancer (SCLC) accounts for about 15-20% of all lung cancer cases and is one of the most malignant tumors. In more than 90% of cases of SCLC, mutations of the TP53 gene are detected. The aim of our study was to analyze the structural changes in the 17p13 chromosomal region in patients with SCLC from the Republic of Bashkortostan. We used 70 lung tissue samples from patients with SCLC. The study was conducted using MLPA. Deletions and/or duplications in the coding regions of the TP53 gene were detected in 40% of the samples, including duplications of the first exon (23% of the samples) and deletions of the 4-6th and 11th exons of the TP53 gene (17% of the samples). Besides, the CHEK2 c.1100delC mutation was detected in four samples.
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Warshawsky, Ilka, Olga B. Chernova, Christian A. Hübner, Reinhard Stindl, Marco Henneke, Andreas Gal i Marvin R. Natowicz. "Multiplex Ligation-Dependent Probe Amplification for Rapid Detection of Proteolipid Protein 1 Gene Duplications and Deletions in Affected Males and Carrier Females with Pelizaeus–Merzbacher Disease". Clinical Chemistry 52, nr 7 (1.07.2006): 1267–75. http://dx.doi.org/10.1373/clinchem.2006.067967.
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Abstract Background: Pelizaeus–Merzbacher disease is a rare X-linked neurodegenerative disorder caused by sequence variations in the proteolipid protein 1 gene (PLP1). PLP1 gene duplications account for ∼50%–75% of cases and point variations for ∼15%–20% of cases; deletions and insertions occur infrequently. We used multiplex ligation-dependent probe amplification (MLPA) to detect PLP1 gene alterations, especially gene duplications and deletions. Methods: We performed MLPA on 102 samples from individuals with diverse PLP1 gene abnormalities and from controls, including 50 samples previously characterized for the PLP1 gene by quantitative PCR but which were anonymized for prior results and sex. Results: All males with PLP1 gene duplications (n = 13), 1 male with a triplication, 2 males with whole gene deletions, and all controls (n = 72) were unambiguously assigned to their correct genotype. Of 4 female carriers tested by MLPA and quantitative PCR, 3 were duplication carriers by both methods, and 1 was a triplication carrier by MLPA and a duplication carrier by quantitative PCR. For 1 sample with a partial deletion, MLPA showed exon 3 deleted but PCR showed exons 3 and 4 deleted. Sequence analysis of 2 samples with reduced dosage for exons 3 and 5 revealed point variations overlapping the annealing site for the corresponding MLPA probe. The precision of MLPA analysis was excellent and comparable to or better than quantitative PCR, with CVs of 4.3%–9.8%. Conclusions: MLPA is a rapid and reliable method to determine PLP1 gene copies. Samples with partial PLP1 gene dosage alterations require confirmation with a non-MLPA method.
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Nichol, Ariadne A., Jason N. Batten, Meghan C. Halley, Julia K. Axelrod, Pamela L. Sankar i Mildred K. Cho. "A Typology of Existing Machine Learning–Based Predictive Analytic Tools Focused on Reducing Costs and Improving Quality in Health Care: Systematic Search and Content Analysis". Journal of Medical Internet Research 23, nr 6 (22.06.2021): e26391. http://dx.doi.org/10.2196/26391.
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Background Considerable effort has been devoted to the development of artificial intelligence, including machine learning–based predictive analytics (MLPA) for use in health care settings. The growth of MLPA could be fueled by payment reforms that hold health care organizations responsible for providing high-quality, cost-effective care. Policy analysts, ethicists, and computer scientists have identified unique ethical and regulatory challenges from the use of MLPA in health care. However, little is known about the types of MLPA health care products available on the market today or their stated goals. Objective This study aims to better characterize available MLPA health care products, identifying and characterizing claims about products recently or currently in use in US health care settings that are marketed as tools to improve health care efficiency by improving quality of care while reducing costs. Methods We conducted systematic database searches of relevant business news and academic research to identify MLPA products for health care efficiency meeting our inclusion and exclusion criteria. We used content analysis to generate MLPA product categories and characterize the organizations marketing the products. Results We identified 106 products and characterized them based on publicly available information in terms of the types of predictions made and the size, type, and clinical training of the leadership of the companies marketing them. We identified 5 categories of predictions made by MLPA products based on publicly available product marketing materials: disease onset and progression, treatment, cost and utilization, admissions and readmissions, and decompensation and adverse events. Conclusions Our findings provide a foundational reference to inform the analysis of specific ethical and regulatory challenges arising from the use of MLPA to improve health care efficiency.
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Fu, Xiaoni, Yinmin Shi, Jiying Ma, Kaiqian Zhang, Guowei Wang, Gang Li, Lei Xiao i Huijuan Wang. "Advances of multiplex ligation-dependent probe amplification technology in molecular diagnostics". BioTechniques 73, nr 4 (październik 2022): 205–13. http://dx.doi.org/10.2144/btn-2022-0017.
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Multiplex ligation-dependent probe amplification (MLPA) is a multiplex copy number analysis tool which is routinely used to detect large mutations in genetic diseases. With continuous modifications, MLPA has been extended for the detection of DNA methylation variation, single nucleotide polymorphisms, chromosome abnormalities and other forms of genomic variation. The combination with other techniques has even enlarged the application of MLPA in molecular diagnostics of various human diseases. In this review, the principle of MLPA-based techniques as well as their main and latest applications in clinical detection are described. It is believed that with improved automation, increased multiplexing, lower cost and the combination with other technologies, MLPA will play an increasingly important role in molecular diagnosis of human disease.
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Zafeiriou, D., A. Ververi, G. Salomons, E. Vargiami, D. Haas, V. Papadopoulou, E. Kontopoulos i C. Jacobs. "MLP05 L-2-hydroxyglutaric aciduria presenting with severe autism". European Journal of Paediatric Neurology 11 (wrzesień 2007): 68. http://dx.doi.org/10.1016/s1090-3798(08)70505-1.
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Coffa, Jordy, Mark A. van de Wiel, Begoña Diosdado, Beatriz Carvalho, Jan Schouten i Gerrit A. Meijer. "MLPAnalyzer: Data Analysis Tool for Reliable Automated Normalization of MLPA Fragment Data". Analytical Cellular Pathology 30, nr 4 (1.01.2008): 323–35. http://dx.doi.org/10.1155/2008/605109.
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Background: Multiplex Ligation dependent Probe Amplification (MLPA) is a rapid, simple, reliable and customized method for detection of copy number changes of individual genes at a high resolution and allows for high throughput analysis. This technique is typically applied for studying specific genes in large sample series. The large amount of data, dissimilarities in PCR efficiency among the different probe amplification products, and sample-to-sample variation pose a challenge to data analysis and interpretation. We therefore set out to develop an MLPA data analysis strategy and tool that is simple to use, while still taking into account the above-mentioned sources of variation.Materials and Methods: MLPAnalyzer was developed in Visual Basic for Applications, and can accept a large number of file formats directly from capillary sequence systems. Sizes of all MLPA probe signals are determined and filtered, quality control steps are performed, and variation in peak intensity related to size is corrected for. DNA copy number ratios of test samples are computed, displayed in a table view and a set of comprehensive figures is generated. To validate this approach, MLPA reactions were performed using a dedicated MLPA mix on 6 different colorectal cancer cell lines. The generated data were normalized using our program and results were compared to previously performed array-CGH results using both statistical methods and visual examination.Results and Discussion: Visual examination of bar graphs and direct ratios for both techniques showed very similar results, while the average Pearson moment correlation over all MLPA probes was found to be 0.42. Our results thus show that automated MLPA data processing following our suggested strategy may be of significant use, especially when handling large MLPA data sets, when samples are of different quality, or interpretation of MLPA electropherograms is too complex. It remains, however, important to recognize that automated MLPA data processing may only be successful when a dedicated experimental setup is also considered.
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Moelans, Cathy B., Roel A. de Weger, Marja T. M. van Blokland, Chantal Ezendam, Sabrina Elshof, Marcel G. J. Tilanus i Paul J. van Diest. "HER-2/neu Amplification Testing in Breast Cancer by Multiplex Ligation-Dependent Probe Amplification in Comparison with Immunohistochemistry and In Situ Hybridization". Analytical Cellular Pathology 31, nr 1 (1.01.2009): 1–10. http://dx.doi.org/10.1155/2009/615178.
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Background: Assessment of HER-2/neu status in invasive breast cancer is crucial to establish eligibility for trastuzumab and taxane based chemotherapy. Next to immunohistochemistry (IHC) to evaluate protein overexpression, a second line gene amplification test is required for cases with equivocal protein expression. This study aimed to validate a new PCR based test, called Multiplex Ligation-dependent Probe Amplification (MLPA), as a simple and quick method to assess HER-2/neu gene amplification status in invasive breast cancer.Methods: MPLA results were compared with gene amplification status assessed by fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) as gold standard, and with protein overexpression by IHC in 518 breast carcinoma patients.Results: About 10% of cases overexpressed HER-2/neu at the protein level (IHC), and 11% of cases showed gene-amplification by MLPA. A high concordance was found between FISH and CISH, MLPA and IHC, and MLPA and CISH. MLPA showed amplification in 7/36 (19%) of the equivocal IHC 2+ cases. However, of the IHC 0/1+ cases, 6/434 (1.4%) were also amplified by MLPA, and amplification was confirmed in all of these cases by FISH/CISH. On the other hand, one of the 48 (2%) IHC 3+ cases was normal by MLPA and lack of amplification was confirmed by FISH/CISH.Conclusion: MLPA is a fast, accurate and cheap method to detect breast cancer HER-2/neu amplification in small quantities of DNA extracted from paraffin blocks, and thereby a reliable alternative to FISH and CISH.
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Razim, Oleksandra, Stefano Cavuoti, Massimo Brescia, Giuseppe Riccio, Mara Salvato i Giuseppe Longo. "Improving the reliability of photometric redshift with machine learning". Monthly Notices of the Royal Astronomical Society 507, nr 4 (13.08.2021): 5034–52. http://dx.doi.org/10.1093/mnras/stab2334.
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ABSTRACT In order to answer the open questions of modern cosmology and galaxy evolution theory, robust algorithms for calculating photometric redshifts (photo-z) for very large samples of galaxies are needed. Correct estimation of the various photo-z algorithms’ performance requires attention to both the performance metrics and the data used for the estimation. In this work, we use the supervised machine learning algorithm MLPQNA (Multi-Layer Perceptron with Quasi-Newton Algorithm) to calculate photometric redshifts for the galaxies in the COSMOS2015 catalogue and the unsupervised Self-Organizing Maps (SOM) to determine the reliability of the resulting estimates. We find that for zspec < 1.2, MLPQNA photo-z predictions are on the same level of quality as spectral energy distribution fitting photo-z. We show that the SOM successfully detects unreliable zspec that cause biases in the estimation of the photo-z algorithms’ performance. Additionally, we use SOM to select the objects with reliable photo-z predictions. Our cleaning procedures allow us to extract the subset of objects for which the quality of the final photo-z catalogues is improved by a factor of 2, compared to the overall statistics.
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Štefeková, Andrea, Pavlína Čapková, Václava Curtisová, Enghjargalan Mracká, Hana Filipová, Zuzana Spurná, Martin Procházka, Marek Ľubušký, Radovan Pilka i Radek Vrtěl. "Prenatal detection of copy number variants in fetuses with detected congenital devolpmental disordes, from 2015 to 2020 by Multiplex Ligation-Dependent Probe Amplification and microarray analysis". Česká gynekologie 88, nr 3 (21.06.2023): 162–71. http://dx.doi.org/10.48095/cccg2023162.
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Objective: Analysis of prenatal samples from 2015 to 2020. Comparison detection rates of clinically relevant variants by cytogenetic karyotype analysis and cytogenomic MLPA (Multiplex Ligation-Depent Probe Amplification) and microarray methods (CMA – chromosomal microarray). Material and method: 1,029 prenatal samples were analyzed by cytogenetic karyotyping (N = 1,029), cytogenomic methods – MLPA (N = 144) and CMA (N = 111). All unbalanced changes were confirmed by MLPA or CMA. Results: From the analyzed set of fetuses, after subtraction of aneuploidies – 107 (10.40%, N = 1,029), 22 structural aberrations (2.39%, N = 922) – nine unbalanced changes (0.98%), 10 balanced changes (1.08%), one case of unclear mosaicism (0.09%), one case of presence of a marker chromosome (0.09%) and one case of sex discordance (0.09%) – were detected by karyotype analysis. A total of eight (7.21%, N = 111) pathological variants were detected by CMA in 255 samples with physiological karyotype indicated for cytogenomic examination. Five (3.47%, N = 144) of eight pathogenic variants were detected by MLPA method. The total capture of pathogenic variants by MLPA and CMA methods was 14 (5.14%) and 17 (6.25%) (N = 272), including confirmatory pathological karyotype testing. Detection of pathological variants in the isolated disorders group was lower than in the multiple disorders group (5.08 vs. 21.42%). Conclusion: A higher success rate for the detection of pathological copy number variation variants by the microarray method than by the MLPA method was confirmed. Key word: congenital developmental disorders – CMA – MLPA – copy number variants
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Moelans, Cathy B., Hanneke N. Monsuur, Johannes H. de Pinth, Remco D. Radersma, Roel A. de Weger i Paul J. van Diest. "ESR1 Amplification is Rare in Breast Cancer and is Associated with High Grade and High Proliferation: A Multiplex Ligation-Dependent Probe Amplification Study". Analytical Cellular Pathology 33, nr 1 (2010): 13–18. http://dx.doi.org/10.1155/2010/619180.
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Background: Expression of estrogen receptor alpha (ERα) is predictive for endocrine therapy response and an important prognostic factor in breast cancer. Overexpression of ERα can be caused by estrogen receptor 1 (ESR1) gene amplification and was originally reported to be a frequent event associated with a significantly longer survival for ER-positive women treated with adjuvant tamoxifen monotherapy, which was however questioned by subsequent studies.Methods: This study aimed to reanalyze the frequency of ESR1 amplification by multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridisation (FISH), and to assess clinicopathologic correlations. MLPA was performed in a group of 135 breast cancer patients, and gains/amplifications were subjected to FISH.Results: True ESR1 amplification by MLPA was rare (2%) and only 6% more patients showed a modest gain of ESR1. All MLPA-detected ESR1 amplifications and nearly all ESR1 gains were also FISH amplified and gained, but not all FISH amplifications/gains were MLPA amplified/gained, leading to an overall concordance of only 60% between both techniques. All 3 MLPA and FISH ESR1 amplified cases had high ERα expression, but there was no obvious correlation between ESR1 gain and ER status by IHC. ESR1 gains/amplifications were not associated with HER2 gain/amplification, but seemed to be associated with older age. Surprisingly, ESR1 gain/amplification was not associated with low grade as reported previously, but correlated with high grade and high proliferation. Furthermore, ESR1 gain/amplification by MLPA was not associated with nodal status or tumor size (pT status).Conclusion: ESR1 amplification as detected by MLPA is rare in breast cancer, and seems to be associated with high ERα expression, high age, high grade and high proliferation. This study confirms previous studies that showed differences in the ESR1 amplification frequencies detected by different techniques.
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Moerland, Elna, Rens L. H. P. M. van Hezik, Toine C. J. M. van der Aa, Mike W. P. M. van Beek i Adriaan J. C. van den Brule. "Detection of HER2 Amplification in Breast Carcinomas: Comparison of Multiplex Ligation-Dependent Probe Amplification (MLPA) and Fluorescence In Situ Hybridization (FISH) combined with Automated Spot Counting". Analytical Cellular Pathology 28, nr 4 (1.01.2006): 151–59. http://dx.doi.org/10.1155/2006/741586.
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In this study the detection of HER2 gene amplification was evaluated using Fluorescence In Situ Hybridization (FISH; PathVysion) in comparison with Multiplex Ligation-dependent Probe Amplification (MLPA), a PCR based technique. These two methods were evaluated on a series of 46 formalin fixed paraffin embedded breast carcinomas, previously tested for protein overexpression by HercepTest (grouped into Hercep 1+, 2+ and 3+). HER2 gene amplification (ratio ≥ 2.0) by FISH was found in 9/10, 10/30 and 0/6 in IHC 3+, 2+ and 1+/0 cases, respectively. Digitalized automated spot counting performed with recently developed CW4000 CytoFISH software was 100% concordant with manual FISH scoring. Using MLPA 18/46 samples showed a clear HER2 amplification. Comparing MLPA and IHC showed the same results as for FISH and IHC. All but one FISH positive cases (18/19) were confirmed by MLPA for the presence of the gene amplification. The overall concordance of detection of Her2 gene amplification by FISH and MLPA was 98% (45/46). Furthermore, both the level of amplification and equivocal results correlated well between both methods. In conclusion, MLPA is a reliable and reproducible technique and can be used as an either alternative or additional test to determine HER2 status in breast carcinomas.
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Abdool, Adam, Amber C. Donahue, Jay G. Wohlgemuth i Chen-Hsiung Yeh. "Detection, Comprehensive Analysis and Clinical Validation of Chromosomal Aberrations by Multiplex Ligation-Dependent Probe Amplification In Chronic Lymphocytic Leukemia". Blood 116, nr 21 (19.11.2010): 2716. http://dx.doi.org/10.1182/blood.v116.21.2716.2716.
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Abstract Abstract 2716 Background: Current strategies based on karyotyping or fluorescent in situ hybridization (FISH) for detection of chromosomal abnormalities in chronic lymphocytic leukemia (CLL) are laborious, time-consuming, and costly and have limitations in resolution. Multiplex ligation-dependent probe amplification (MLPA) can simultaneously detect copy number changes of multiple loci in a single PCR, making it an attractive alternative. We developed and validated an MLPA protocol for comprehensive, automated data analysis and interpretation of chromosome abnormalities associated with CLL. Patients and Methods: Reference ranges for individual MLPA probes were established from a group of 50 healthy control subjects. Using these ranges we built an automated spreadsheet-based analysis system that includes multiple quality checks; samples that fail these checks are flagged and not reported. Each target was given a call of “deletion,” “normal,” or “amplification,” depending on whether the normalized ratio fell within or outside of the established normal range (mean ± 2SD or mean ± 3SD). After establishing the normal references ranges for each probe, we used the MLPA assay to characterize chromosome abnormalities in blood samples from 100 patients with suspected CLL that had been previously tested with FISH. Results: The maximum normal ranges were distributed between 0.82 and 1.18 for the mean ± 2SD values (ie, 95% CI, P = 0.05), and between 0.73 and 1.27 for the mean ± 3SD values (ie, 99.7% CI, P = 0.01). MLPA showed good concordance with FISH results in the 100 clinically suspected CLL cases. In 6 of these cases, abnormalities detected by MLPA were in regions not covered by FISH, including additional copy number gains on chromosomes 18q21.1 and 19, and novel micro-deletions at 19q13.43 and 19p13.2 loci. Excluding these cases, MLPA showed 94% sensitivity, 94% concordance, and 93% specificity relative to FISH. MLPA detected abnormalities in 3 FISH-negative cases and failed to detect abnormalities in three 13q- cases with low percentages of leukemic cells (7%, 12% and 19%). The limit of detection of the CLL MLPA assay was about 20% leukemic clones. Conclusions: This MLPA-based assay for chromosome abnormalities in CLL showed excellent concordance with FISH. This multiplex assay represents a fast (roughly 2–3 days total process to report time vs 7–10 days for FISH), high-throughput, accurate, and user-friendly process for that has potential for use as a first-line screening test for detection of chromosome abnormalities associated with CLL in the clinical laboratory. Disclosures: No relevant conflicts of interest to declare.
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Pirelli, Kevin, Curt Czajkowski, Veronika Mladenova, Stefanie Dugan, Valerie Trapp-Stamborski, Kenneth D. Friedman i Rupa A. Udani. "Laboratory-Developed MLPA® Probes Enable CNV Detection in the CFHR4 Gene to Aid aHUS Genetic Evaluation". Blood 128, nr 22 (2.12.2016): 1361. http://dx.doi.org/10.1182/blood.v128.22.1361.1361.
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Abstract Large copy number variants (CNVs) such as deletions in the regulator of complement activation (RCA) gene cluster (CFH, CFHR3, CFHR1, CFHR4, CFHR2, CFHR5) are associated with elevated risk for atypical hemolytic uremic syndrome (aHUS), a complement regulation disorder (Zipfel 2007). Of particular interest are compound heterozygous CFHR3-CFHR1 and CFHR1-CFHR4 deletions, linked to aHUS due to the resulting composite homozygous CFHR1 deletions (Moore 2010). Detection of CFHR4 CNVs is important as it enables identification of said CFHR3-CFHR1 deletion in trans with the CFHR1-CFHR4 deletion, as well as rare reported CFHR4 duplications (Sudmant 2010). Large heterozygous deletions or duplications involving CFHR3 and CFHR1 or CFHR1 and CFHR4 are undetectable by Sanger sequencing and difficult to detect with amplicon-based Next-Generation Sequencing (NGS). When CNVs are detected by NGS, confirmation with an independent reference method is required by regulatory agencies including the New York State Department of Health and College of American Pathology. Multiplex Ligation-Dependent Probe Amplification (MLPA®) has been used to reliably detect large CNVs for CFH, CFHR3, CFHR1, CFHR2 and CFHR5, and a commercial research use only (RUO) probe mix is available for this purpose from MRC-Holland (Amsterdam, NL). For the CFHR4 region, however, MLPA® probes are not available from any commercial source. Furthermore, CFHR4 MLPA® probe design is complicated by high internal and external homology with adjacent RCA regions (including 91.3-96.2% homology within CFHR4 exons, as well as 90-100% homology with CFHR3 exons). Using a combination of MLPA® probe design freeware, commercial software, literature sources and in-house modifications, a multiplex MLPA® laboratory-developed test (LDT) targeting 5 regions of interest (ROIs) in the CFHR4 gene was designed and validated. The multiplex CFHR4 MLPA® LDT was customized to include an internally-designed ROI MLPA® probe mix targeting CFHR4 intron 1, exon 2, exon 5, intron 6, intron 9, as well as commercially available MLPA® quality control and reference probes. Exon 2 probe sequences were obtained from the literature (Kubista 2011). The remaining MLPA® ROI probes were designed using AlleleID® software (Premier Biosoft) or online freeware MAPD-MLPA (courtesy Stony Brook University Bioinformatics). The MLPA®CFHR4 ROI probes were optimized and challenged to achieve complete specificity for and coverage across the CFHR4 region, as well as to prevent overlap with quality control and reference probes. The CFHR4 MLPA® LDT was validated with 10 genomic DNA samples, comprising a mixture of healthy donors or patients (BloodCenter of Wisconsin archives) and 6 samples from Coriell Repositories (Camden, NJ), all of which were known to harbor large RCA CNVs. 100% concordance with expected findings was observed. Full aHUS genetic evaluation comprising NGS analysis of 15 genes and MLPA® analysis of the RCA gene cluster has now been performed for 161 patients. In 95 (59%) of these patients, no large CNV was detected. Forty patients (25%) tested negative for any large CFHR4 variant but did test positive for the CFHR3-CFHR1 heterozygous deletion. 28% of the population are reported heterozygous for this CFHR3-CFHR1 deletion (Moore 2010). Fifteen patients (9%) were positive for homozygous CFHR1 deletions, which are associated with increased risk for development of autoantibody to complement factor H (CFH), as reported by Moore 2010. Using the CFHR4 MLPA® LDT, the latter homozygous CFHR1 deletions were attributed to compound heterozygosity for the CFHR3-CFHR1 and CFHR1-CFHR4 deletions in 3 of the aforesaid 15 patients. The other 12 patients were found to harbor homozygous deletions of CFHR3 and CFHR1. In 3 (1.9%) patients, the CFHR4 MLPA® LDT revealed a CFHR1-CFHR4 heterozygous deletion, similar to the previously reported frequency of 1.4% in aHUS patients (Moore 2010). Finally, 2 patients tested positive for a heterozygous CFHR3 deletion and a CFHR4 duplication, consistent with the presence of a CFHR3-CFHR1 deletion on one allele and CFHR1-CFHR4 duplication on the other allele resulting in a normal CFHR1 copy number. Overall, these data show the importance of CFHR4 MLPA® CNV testing to aid genetic evaluation of suspected aHUS cases, and this testing should be part of the aHUS genetic panel. Disclosures Friedman: Alexion: Speakers Bureau; NovoNordisk: Consultancy; Shire: Consultancy; CSL Behring: Consultancy.
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Sharma, Pankaj, Neerja Gupta, Madhumita R. Chowdhury, Shubha R. Phadke, Savita Sapra, Ashutosh Halder, Manju Ghosh i Madhulika Kabra. "Williams-Beuren Syndrome: Experience of 43 Patients and a Report of an Atypical Case from a Tertiary Care Center in India". Cytogenetic and Genome Research 146, nr 3 (2015): 187–94. http://dx.doi.org/10.1159/000439205.
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Williams-Beuren syndrome (WBS) or Williams syndrome (OMIM 194050) is a multisystem disorder manifested by neurodevelopmental delay and is caused by a hemizygous deletion of ∼1.5-1.8 Mb in the 7q11.23 region. Clinical features include cardiovascular anomalies (mainly supravalvular aortic stenosis), peripheral pulmonary stenosis, distinctive facies, intellectual disability (usually mild), unique personality characteristics, and growth and endocrine abnormalities. Clinical diagnostic criteria are available for WBS; however, the mainstay of diagnosis is the detection of the contiguous gene deletion. Although FISH remains the most widely used laboratory test, the diagnosis can also be established by means of qPCR, MLPA, microsatellite marker analysis, and chromosomal microarray (CMA). We evaluated the utility of MLPA to detect deletion/duplication in the 7q11.23 region in 43 patients suspected to have WBS using MLPA kits for microdeletion syndromes. A hemizygous deletion in the 7q11.23 region was found in 41 (95.3%) patients using MLPA. One patient had an atypical deletion detected by CMA. During the initial period of this study, the results of 12 patients tested by MLPA were also confirmed by FISH. Compared to FISH and CMA, MLPA is a cheaper, high-throughput, less labor-intensive and less time-consuming technique for the diagnosis of WBS. Although CMA is expensive and labor-intensive, its effectiveness is demonstrated to detect an atypical deletion and to delineate the breakpoints.
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Zagradišnik, Boris, Špela Stangler Herodež, Alenka Erjavec-Škerget, Andreja Zagorac i Nadja Kokalj-Vokač. "Detection of aneuploidy using multiplex ligation–dependent probe amplification in fetal tissues from aborted pregnancies". Acta Medico-Biotechnica 4, nr 2 (27.11.2021): 51–60. http://dx.doi.org/10.18690/actabiomed.58.
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Purpose: About 10-15% of all pregnancies terminate as spontaneous miscarriages. In the first trimester, ≈50% of spontaneous miscarriages are the result of chromosomal aberrations, mostly chromosomal aneuploidies. Cytogenetic analyses are used to confirm aneuploidy in failed pregnancies. Culture failure or poor-quality chromosomes are often problems in those cases. In such situations, methods that are independent of tissue culture are used, and we employed multiplex ligation-dependent probe amplification (MLPA). We determined if MLPA is an appropriate and compatible method compared with classical cytogenetic analyses on fetal tissues.
Methods: All fetal samples received from spontaneous abortions were cultured, karyotyped (if possible) and genomic DNA extracted. MLPA analyses were undertaken using subtelomeric probe kits. Additionally, comparative genomic hybridization (CGH) was used to confirm aneuploidy detected by MLPA in cases of failed culture growth.
Results: MLPA analyses confirmed an unbalanced chromosome abnormality identified by cytogenetic analyses in all cases in which tissue culture was successful, and provided data in cases of failed culture growth. Several common numeric chromosome aberrations were detected, as well as rare trisomies and other unbalanced chromosome rearrangements.
Conclusions: MLPA analyses can provide information about the karyotype of a DNA sample if cytogenetic analyses are not possible because of a lack of viable cells or if only a small amount of genomic DNA is available. Th
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Izadmanesh, Yahya, Jahan B. Ghasemi i Roma Tauler. "Receptor modeling of environmental aerosol data using MLPCA-MCR-ALS". Chemometrics and Intelligent Laboratory Systems 167 (sierpień 2017): 50–62. http://dx.doi.org/10.1016/j.chemolab.2017.05.008.
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Cavuoti, S., M. Brescia, G. Longo i A. Mercurio. "Photometric redshifts with the quasi Newton algorithm (MLPQNA) Results in the PHAT1 contest". Astronomy & Astrophysics 546 (28.09.2012): A13. http://dx.doi.org/10.1051/0004-6361/201219755.
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Nhung, Vu Phuong, Nguyen Thi Thanh Hoa, Ma Thi Huyen Thuong, Tran Thi Bich Ngoc, Nguyen Dang Ton, Nguyen Thuy Duong, Nong Van Hai i Nguyen Hai Ha. "STUDY ON APPLICATION OF MULTIPLEX LIGATION-DEPENDENT PROBE AMPLIFICATION (MLPA) ASSAY IN MOLECULAR DIAGNOSIS OF RETINOBLASTOMA". Vietnam Journal of Biotechnology 15, nr 4 (14.12.2018): 625–31. http://dx.doi.org/10.15625/1811-4989/15/4/13402.
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Retinoblastoma (Rb) is a malignant retinal tumor on young children which is often founded before the age of 5. This cancer disease appears when both of RB1 alleles on 13q14.2 chromosome are mutated. The aim of this research is to evaluate the ability of Multiplex Ligation-dependent Probe Amplification (MLPA) in screening of RB1 gene insertion/deletions in Vietnamese patients with Rb. Genomic DNA was isolated from peripheral blood of the research subjects and subsequently analyzed by MLPA technique. To prove the results of MLPA, quantitative real-time PCR was used for determining the RB1 gene copy number for all samples. Two significant deletion mutations were identified on two different Rb patients, one is the deletion from exon 4 to exon 27 recorded on KVM38 sample, and the other is the complete removal of an allele on KVM21 sample. The MLPA showed a complete correlation with real-time PCR results. These are the disease causing mutations, can be inherited and they are important evidences for genetic counseling and clinical management. Those results have proven the high speed and reliability of MLPA method in identifying deletion/duplication mutations on Rb patients.
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Donahue, Amber C., Adam Abdool, Jay G. Wohlgemuth i Chen-Hsiung Yeh. "Molecular Characterization of Chromosomal Abnormalities In Myelodysplastic Syndrome and Acute Myeloid Leukemia: Validation of An MLPA Protocol and Analysis Method for Use In a Diagnostic Setting". Blood 116, nr 21 (19.11.2010): 4849. http://dx.doi.org/10.1182/blood.v116.21.4849.4849.
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Abstract Abstract 4849 Introduction: Current diagnostic screening strategies for copy number variations (CNVs) in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) include fluorescence in situ hybridization (FISH) or karyotyping, both of which are time-consuming, costly, laborious, and lacking in resolution. Multiplex ligation-dependent probe amplification (MLPA) can be used to detect copy number changes in multiple loci simultaneously in a single PCR reaction, and boasts a resolution down to single exons. To adapt MLPA for use in routine clinical diagnostics, we have developed and validated a protocol for automatic data analysis and interpretation of common chromosomal abnormalities in MDS/AML. Patients and Methods: The study used a training set of 45 healthy subjects to establish a normal reference range for each individual probe. Using these ranges we built an automated Excel spreadsheet-based analysis system, which included multiple quality checks, and flagged samples failing these quality controls. Each probe was given a call of “no mutation detected,” “deletion,” or “gain,” based on whether the normalized ratio fell within or outside of the empirically-determined normal range for that probe. We then analyzed over 100 leukemia cases tested by FISH, including both suspected myeloid leukemia samples and suspected chronic lymphocytic leukemia (CLL) samples. Documented chromosomal abnormalities in CLL include 11q-, 17p- (loss of TP53), and trisomy 12, all of which had the potential to be detected by the probes in the MDS MLPA probemix. The greater prevalence of CLL and its associated CNVs provided additional positive controls for the validation of the MDS MLPA probemix and our analysis method. Results: The empirically-determined normal ranges demonstrated that some probes varied widely (3 standard deviation [3SD] normal range of 0.46–1.54), while others were extremely reliable (3SD normal range of 0.84–1.16). The MLPA assay demonstrated excellent overall accuracy (>90%) and specificity (>93%) for both suspected myeloid and CLL samples when compared to FISH. The sensitivity of the MLPA assay is somewhat lower than that of FISH, requiring a probe-dependent 20–40% positivity for a given CNV to be detected. However in several cases, the MDS MLPA assay was able to detect additional lesions too small to be seen by FISH. Conclusions: For MLPA, the total process-to-report time, including data analysis, is 2–3 days, versus the 7–10 days required for FISH analysis. In addition, the MLPA assay is substantially cheaper and considerably less labor-intensive than FISH. Our improved MLPA assay protocol and analysis method provides a clinically robust, multiplexed, high-throughput, high-resolution, and low-cost solution for detection of copy number changes in MDS/AML, and can therefore be used as a first-line screening test in a clinical laboratory. Disclosures: Donahue: Quest Diagnostics Inc.: Employment. Abdool: Quest Diagnostics Inc.: Employment. Wohlgemuth: Quest Diagnostics Inc.: Employment. Yeh: Quest Diagnostics Inc.: Employment.
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Funari, Mariana F. A., Alexander A. L. Jorge, Emilia M. Pinto, Ivo J. P. Arnhold, Berenice B. Mendonca i Mirian Y. Nishi. "Cryptic intragenic deletion of the SHOX gene in a family with Léri-Weill dyschondrosteosis detected by Multiplex Ligation-Dependent Probe Amplification (MLPA)". Arquivos Brasileiros de Endocrinologia & Metabologia 52, nr 8 (listopad 2008): 1382–87. http://dx.doi.org/10.1590/s0004-27302008000800029.
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LWD is associated to SHOX haploinsufficiency, in most cases, due to gene deletion. Generally FISH and microsatellite analysis are used to identify SHOX deletion. MLPA is a new method of detecting gene copy variation, allowing simultaneous analysis of several regions. Here we describe the presence of a SHOX intragenic deletion in a family with LWD, analyzed through different methodologies. Genomic DNA of 11 subjects from one family were studied by microsatellite analysis, direct sequencing and MLPA. FISH was performed in two affected individuals. Microsatellite analysis showed that all affected members shared the same haplotype suggesting the involvement of SHOX. MLPA detected an intragenic deletion involving exons IV-VIa, which was not detected by FISH and microsatellite analysis. In conclusion, the MLPA technique was proved to be the best solution on detecting this small deletion, it has the advantage of being less laborious also allowing the analysis of several regions simultaneously.
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Schmid, Andreas, Martin Huonker, Jose-Miguel Barturen, Fabian Stahl, Arno Schmidt-Trucksäss, Daniel König, Dominik Grathwohl, Manfred Lehmann i Joseph Keul. "Catecholamines, heart rate, and oxygen uptake during exercise in persons with spinal cord injury". Journal of Applied Physiology 85, nr 2 (1.08.1998): 635–41. http://dx.doi.org/10.1152/jappl.1998.85.2.635.
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The purpose of this study was to investigate the influence of different injury levels in persons with spinal cord injury (SCI) on epinephrine (Epi) and norepinephrine (NE) at rest and during graded wheelchair exercise and the related changes in heart rate and O2 uptake (V˙o 2). Twenty tetraplegics (Tetra), 10 high-lesion paraplegics (HLPara), 20 paraplegics with SCI below T5 (MLPara), and 18 able-bodied, nonhandicapped persons (AB) were examined. Because of the higher level of interruption of the sympathetic pathways, Tetra persons showed lower Epi and NE at rest and only slight increases during exercise compared with all other groups; the Tetra subjects’ impaired cardiac sympathetic innervation caused restricted cardioacceleration and strongly reduced maximalV˙o 2. When compared with AB persons, HLPara had comparable NE but lower Epi levels as a result of partial innervation of the noradrenergic system and denervation of the adrenal medulla. MLPara subjects showed an augmented basal and exercise-induced upper spinal thoracic sympathetic activity compared with AB subjects. The increase in heart rate in relation toV˙o 2 was higher in HLPara because of a smaller stroke volume as a result of venous blood pooling. The different exercise response in persons with SCI is a result of the interruption of pathways in the spinal cord to the peripheral sympathetic nervous system in addition to the motor paralysis.
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Barzaghi, C., A. Giovanetti, C. Reale, J. Sequeiros, N. Nardocci, A. Albanese i B. Garavaglia. "P2.062 MLPA analysis in EOP patients". Parkinsonism & Related Disorders 15 (grudzień 2009): S105. http://dx.doi.org/10.1016/s1353-8020(09)70413-4.
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Honjo, Rachel Sayuri, Roberta Lelis Dutra, Erika Arai Furusawa, Evelin Aline Zanardo, Larissa Sampaio de Athayde Costa, Leslie Domenici Kulikowski, Debora Romeo Bertola i Chong Ae Kim. "Williams-Beuren Syndrome: A Clinical Study of 55 Brazilian Patients and the Diagnostic Use of MLPA". BioMed Research International 2015 (2015): 1–6. http://dx.doi.org/10.1155/2015/903175.
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Williams-Beuren syndrome (WBS) is a genetic disease caused by a microdeletion in the 7q11.23 region. It is characterized by congenital heart disease, mainly supravalvular aortic stenosis, mental retardation, mild short stature, facial dysmorphisms, and variable abnormalities in different systems.Objectives. To report the clinical findings of 55 Brazilian patients confirmed by multiplex ligation-dependent probe amplification (MLPA).Methods. Patients were followed up for 4 years at the Genetics Unit of the Instituto da Criança of the Hospital das Clínicas, FMUSP, Brazil. A kit specific for WBS was used to detect the 7q11.23 microdeletion.Results. Two patients with negative FISH results had positive MLPA results for WBS. The characteristics of the patients with the deletion were as follows: typical WBS facies (98.2%), neuropsychomotor delay (98.2%), hypersocial behavior (94.5%), hyperacusis (94.5%), and congenital heart disease (81.8%).Conclusions. MLPA was effective in detecting the microdeletion in the 7q11.23 region to confirm the diagnosis of WBS. MLPA was also able to confirm the diagnosis of WBS in two patients with typical clinical characteristics but negative FISH results. Thus, MLPA is a promising method in the diagnostic investigation of WBS. WBS is a multisystemic disorder and therefore requires multidisciplinary care and specific follow-up to prevent complications.
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Aktürk, A., E. Arhan, T. Eminoğlu, G. Biberoğlu, A. Serdaroğlu, L. Tümer, A. Hasanoğlu, E. Demir i K. Gücüyener. "MLP01 3-Methylcrotonyl-CoA carboxylase deficiency: phenotypic variability in a family". European Journal of Paediatric Neurology 11 (wrzesień 2007): 67. http://dx.doi.org/10.1016/s1090-3798(08)70501-4.
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Langius, F., N. Abeling, M. Duran i B. T. Poll-The. "MLP08 Developmental delay and cerebral palsy associated with tryptophan hydroxylase deficiency". European Journal of Paediatric Neurology 11 (wrzesień 2007): 69. http://dx.doi.org/10.1016/s1090-3798(08)70508-7.
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Mäkikangas, Anne, Asko Tolvanen, Kaisa Aunola, Taru Feldt, Saija Mauno i Ulla Kinnunen. "Multilevel Latent Profile Analysis With Covariates". Organizational Research Methods 21, nr 4 (22.02.2018): 931–54. http://dx.doi.org/10.1177/1094428118760690.
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Latent profile analysis (LPA) is a person-centered method commonly used in organizational research to identify homogeneous subpopulations of employees within a heterogeneous population. However, in the case of nested data structures, such as employees nested in work departments, multilevel techniques are needed. Multilevel LPA (MLPA) enables adequate modeling of subpopulations in hierarchical data sets. MLPA enables investigation of variability in the proportions of Level 1 profiles across Level 2 units, and of Level 2 latent classes based on the proportions of Level 1 latent profiles and Level 1 ratings, and the extent to which covariates drawn from the different hierarchical levels of the data affect the probability of a membership of a particular profile. We demonstrate the use of MLPA by investigating job characteristics profiles based on the job-demand-control-support (JDCS) model using data from 1,958 university employees clustered in 78 work departments. The implications of the results for organizational research are discussed, together with several issues related to the potential of MLPA for wider application.
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Sarasati, Shinta Andi, Kristy Iskandar, Maria Alethea Septianastiti, Rusdy Ghazali Malueka i Ery Kus Dwianingsih. "Diagnostic Value of Dystrophin Immunostaining in the Diagnosis of Duchenne and Becker Muscular Dystrophy Patients". Open Access Macedonian Journal of Medical Sciences 9, A (1.12.2021): 1137–41. http://dx.doi.org/10.3889/oamjms.2021.7612.
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Background: Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are X-linked recessive muscular disorders caused by the absence or reduction of the muscle cytoskeletal protein dystrophin. Standard procedures to detect deletion and duplication of the DMD gene use Multiplex Ligation-Dependent Probe Amplification (MLPA). However, genetic testing, such as MLPA, is not covered by the national insurance scheme in Indonesia. Immunohistochemical (IHC) staining of dystrophin from muscle biopsy in the form of Formalin-Fixed Paraffin-Embedded (FFPE) specimens can be an alternative method to detect dystrophin expression in protein levels to establish the diagnosis of DMD or BMD.
Objectives: To determinate sensitivity, specificity and accuracy of IHC analysis of dystrophin in DMD/BMD patient in comparison with the standard genetic testing, MLPA.
Methods: Twenty-six patients enrolled in this study were clinically diagnosed as DMD/BMD in Dr. Sardjito Hospital and Universitas Gadjah Mada Academic Hospital. Genomic DNA was isolated from 3 mL of EDTA-peripheral whole blood samples. The deletion and duplication of DMD genes were detected by MLPA. IHC examination was performed using a specific antibody dystrophin (DYS2). Complete loss of dystrophin staining indicated DMD, while partial loss of dystrophin staining indicated BMD. MLPA result was used as the gold standard to determine sensitivity, specificity, and accuracy of IHC technique using a 2x2 table.
Results: MLPA results revealed 18 (18/26; 69.3%) patients with deletion and 3 (3/26; 11.5%) patients with duplication. Five (5/26; 19.2%) patients who showed no deletion nor duplication were excluded from the analysis. Among 21 patients with deletion or duplication, 18 (18/21; 85.7%) patients were out-of-frame (DMD) and 3 (3/21; 14.3%) patients were in-frame (BMD). Six patients showed a discrepancy between the IHC and MLPA results with 9.5% (2/21) false positive and 19% (4/21) false negative. The sensitivity of dystrophin IHC was 77.78%, specificity 33.33%, positive predictive value 87.5%, negative predictive value 20%, and accuracy 71.43%.
Conclusion: Muscle biopsy followed by IHC can be one of the diagnostic tools to diagnose BMD or DMD, with high sensitivity. The protein-based strategy is probably the most efficient way to approach the diagnosis of Duchenne and Becker muscular dystrophy in limited health care settings.
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Uscanga Morales, Luis Alejandro, i María Perevochtchikova. "De Pago por Servicios Ambientales Hidrológicos a Fondos Concurrentes: estudio de percepción social en una comunidad forestal de Oaxaca, México". Sociedad y Ambiente, nr 23 (28.08.2020): 1–31. http://dx.doi.org/10.31840/sya.vi23.2161.
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Esta investigación contribuye a la discusión sobre el futuro del programa federal de Pago por Servicios Ambientales Hidrológicos (PSAH) y su posible transición hacia los Mecanismos Locales de Pago por Servicios Ambientales a través de Fondos Concurrentes (MLPSA-FC). Se desarrolló un análisis de percepción social con actores clave involucrados en el proceso de aplicación (proveedores, usuarios e intermediarios) sobre los efectos sociales, económicos y ambientales generados en una comunidad forestal de Oaxaca, México. A partir de entrevistas semiestructuradas aplicadas en 2016-2017, se identificó que los usuarios se inclinan hacia los MLPSA-FC; no obstante, los intermediarios y los proveedores prefieren el PSAH. Al parecer, la transición no se da por una decisión voluntaria, sino en un contexto de alta competitividad e insuficiencia presupuestaria para sustentar el programa federal de PSAH por parte de la Comisión Nacional Forestal (CONAFOR). En este sentido, es necesario elaborar evaluaciones de los efectos, desde un enfoque integral, de los esquemas de PSA en México, así como estudios de comparación de los efectos generados por los programas federales y los mecanismos locales desde perspectivas inter y transdisciplinarias.
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Fonseca, R. L., A. R. Lobo-Jr i M. I. S. Santana. "Measurements of femoral angles, femur length, and hip width in cat radiographs". Arquivo Brasileiro de Medicina Veterinária e Zootecnia 69, nr 6 (listopad 2017): 1513–20. http://dx.doi.org/10.1590/1678-4162-9583.
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ABSTRACT Femoral angle, femur length, and hip width were measured in radiographs of 92 intact domestic cats, males and females of mixed breed from the Center for Zoonosis Control of the Federal District. The animals showed no trauma, orthopedic diseases or angular deformities and had closed physeal lines. Accordingly, we measured aLPFA (anatomical lateral proximal femoral angle, aLDFA (anatomical lateral distal femoral angle), mLPFA (mechanical lateral proximal femoral angle), mLDFA (mechanical lateral distal femoral angle), IA (femoral inclination angle), FL (femur length) and HW (hip width) using ventrodorsal radiographs, with both hindlimbs in a single exposure to an X-ray beam centered on the hip. The mean values of the variables were: mLPFA: 82.5±3.62°; aLPFA: 80.1±4.29°; mLDFA: 96.1±3.51° (males) and 97.3±2.05° (females); aLDFA: 94,3±3.43°; IA: 136.6±3.86°; FL: 12.9±0.55cm (males) and 13.4±0.66cm (females); and HW: 3.1cm±0.23 (males) and 3.5±0.26cm (females). These values will serve as a reference for the diagnosis of angular deformities and as support for planning corrective osteotomies in domestic cats.
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Minaidou, Anna, Stella Tamana, Coralea Stephanou, Maria Xenophontos, Cornelis L. Harteveld, Celeste Bento, Marina Kleanthous i Petros Kountouris. "A Novel Tool for the Analysis and Detection of Copy Number Variants Associated with Haemoglobinopathies". International Journal of Molecular Sciences 23, nr 24 (14.12.2022): 15920. http://dx.doi.org/10.3390/ijms232415920.
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Several types of haemoglobinopathies are caused by copy number variants (CNVs). While diagnosis is often based on haematological and biochemical parameters, a definitive diagnosis requires molecular DNA analysis. In some cases, the molecular characterisation of large deletions/duplications is challenging and inconclusive and often requires the use of specific diagnostic procedures, such as multiplex ligation-dependent probe amplification (MLPA). Herein, we collected and comprehensively analysed all known CNVs associated with haemoglobinopathies. The dataset of 291 CNVs was retrieved from the IthaGenes database and was further manually annotated to specify genomic locations, breakpoints and MLPA probes relevant for each CNV. We developed IthaCNVs, a publicly available and easy-to-use online tool that can facilitate the diagnosis of rare and diagnostically challenging haemoglobinopathy cases attributed to CNVs. Importantly, it facilitates the filtering of available entries based on the type of breakpoint information, on specific chromosomal and locus positions, on MLPA probes, and on affected gene(s). IthaCNVs brings together manually curated information about CNV genomic locations, functional effects, and information that can facilitate CNV characterisation through MLPA. It can help laboratory staff and clinicians confirm suspected diagnosis of CNVs based on molecular DNA screening and analysis.
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Gao, Dawei, Bufan Yao, Gaoshuang Chang i Qiang Li. "Multi-Objective Optimization Design of Vehicle Side Crashworthiness Based on Machine Learning Point-Adding Method". Applied Sciences 12, nr 20 (13.10.2022): 10320. http://dx.doi.org/10.3390/app122010320.
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Multi-objective optimization problems are often accompanied by complex black-box functions which not only increases the difficulty of solving, but also increases the solving time. In order to reduce the computational cost of solving such multi-objective problems, this paper proposes an ARBF-MLPA (Adaptive Radial Basis Function neural network combined with Machine Learning Point Adding) method, which uses an ABRF (Adaptive Radial Basis Function) neural network and OLHS (Optimized Latin Hypercube Sampling) to establish the first generation metamodel and uses the NSGA-II (Non-dominated Sorting Genetic Algorithm II) optimization algorithm to obtain the optimal front edge of Pareto. The ARBF-MLPA method is continuously used to select and add points to update the meta-model, then dynamically improve the accuracy of the meta-model until the optimal front edge converges. Then the ARBF-MLPA method and RBF-UDPA (Radial Basis Function neural network combined with Uniform Point Adding) method are compared using the test functions of three different frontier features. The performance evaluation indexes of Inverted Generation Distance (IGD), Hypervolume (HV) and Spacing Metric are superior to RBF-UDPA. Finally, ARBF-MLPA method combined with the NSGA-II optimization algorithm is applied in the multi-objective optimization design of vehicle-side crashworthiness. The model converges after 6 iterations. Comparing the results obtained by the ARBF-MLPA method with the finite element simulation results, the error is within 5%, which meets the error requirements. The optimized model reduces chest intrusion by 4.32%, peak collision force by 2.11% and reduces mass by 14.05%.
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Capkova, Pavlina, Josef Srovnal, Zuzana Capkova, Katerina Staffova, Vera Becvarova, Marie Trkova, Katerina Adamova i in. "MLPA is a practical and complementary alternative to CMA for diagnostic testing in patients with autism spectrum disorders and identifying new candidate CNVs associated with autism". PeerJ 6 (9.01.2019): e6183. http://dx.doi.org/10.7717/peerj.6183.
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Background Autism spectrum disorder (ASD) is a complex heterogeneous developmental disease with a significant genetic background that is frequently caused by rare copy number variants (CNVs). Microarray-based whole-genome approaches for CNV detection are widely accepted. However, the clinical significance of most CNV is poorly understood, so results obtained using such methods are sometimes ambiguous. We therefore evaluated a targeted approach based on multiplex ligation-dependent probe amplification (MLPA) using selected probemixes to detect clinically relevant variants for diagnostic testing of ASD patients. We compare the reliability and efficiency of this test to those of chromosomal microarray analysis (CMA) and other tests available to our laboratory. In addition, we identify new candidate genes for ASD identified in a cohort of ASD-diagnosed patients. Method We describe the use of MLPA, CMA, and karyotyping to detect CNV in 92 ASD patients and evaluate their clinical significance. Result Pathogenic and likely pathogenic mutations were identified by CMA in eight (8.07% of the studied cohort) and 12 (13.04%) ASD patients, respectively, and in eight (8.07%) and four (4.35%) patients, respectively, by MLPA. The detected mutations include the 22q13.3 deletion, which was attributed to ring chromosome 22 formation based on karyotyping. CMA revealed a total of 91 rare CNV in 55 patients: eight pathogenic, 15 designated variants of unknown significance (VOUS)—likely pathogenic, 10 VOUS—uncertain, and 58 VOUS—likely benign or benign. MLPA revealed 18 CNV in 18 individuals: eight pathogenic, four designated as VOUS—likely pathogenic, and six designated as VOUS—likely benign/benign. Rare CNVs were detected in 17 (58.62%) out of 29 females and 38 (60.32%) out of 63 males in the cohort. Two genes, DOCK8 and PARK2, were found to be overlapped by CNV designated pathogenic, VOUS—likely pathogenic, or VOUS—uncertain in multiple patients. Moreover, the studied ASD cohort exhibited significant (p < 0.05) enrichment of duplications encompassing DOCK8. Conclusion Multiplex ligation-dependent probe amplification and CMA yielded concordant results for 12 patients bearing CNV designated pathogenic or VOUS—likely pathogenic. Unambiguous diagnoses were achieved for eight patients (corresponding to 8.7% of the total studied population) by both MLPA and CMA, for one (1.09%) patient by karyotyping, and for one (1.09%) patient by FRAXA testing. MLPA and CMA thus achieved identical reliability with respect to clinically relevant findings. As such, MLPA could be useful as a fast and inexpensive test in patients with syndromic autism. The detection rate of potentially pathogenic variants (VOUS—likely pathogenic) achieved by CMA was higher than that for MLPA (13.04% vs. 4.35%). However, there was no corresponding difference in the rate of unambiguous diagnoses of ASD patients. In addition, the results obtained suggest that DOCK8 may play a role in the etiology of ASD.
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Milunsky, J. M., T. A. Maher, M. Ito i A. Milunsky. "The Value of MLPA in Waardenburg Syndrome". Genetic Testing 11, nr 2 (czerwiec 2007): 179–82. http://dx.doi.org/10.1089/gte.2006.0531.
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ACQUILA, M., M. PASINO, M. DI DUCA, F. BOTTINI, A. C. MOLINARI i M. P. BICOCCHI. "MLPA assay in F8 gene mutation screening". Haemophilia 14, nr 3 (maj 2008): 625–27. http://dx.doi.org/10.1111/j.1365-2516.2008.01659.x.
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