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Littérature scientifique sur le sujet « QLAMP »

Auteur : Grafiati

Publié le 9 mars 2023

Mis à jour le 10 mars 2023

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Articles de revues sur le sujet "QLAMP"

Thiessen, Lindsey D., Tara M. Neill et Walter F. Mahaffee. « Development of a quantitative loop-mediated isothermal amplification assay for the field detection ofErysiphe necator ». PeerJ 6 (20 avril 2018) : e4639. http://dx.doi.org/10.7717/peerj.4639.

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Plant pathogen detection systems have been useful tools to monitor inoculum presence and initiate management schedules. More recently, a loop-mediated isothermal amplification (LAMP) assay was successfully designed for field use in the grape powdery mildew pathosystem; however, false negatives or false positives were prevalent in grower-conducted assays due to the difficulty in perceiving the magnesium pyrophosphate precipitate at low DNA concentrations. A quantitative LAMP (qLAMP) assay using a fluorescence resonance energy transfer-based probe was assessed by grape growers in the Willamette Valley of Oregon. Custom impaction spore samplers were placed at a research vineyard and six commercial vineyard locations, and were tested bi-weekly by the lab and by growers. Grower-conducted qLAMP assays used a beta-version of the Smart-DART handheld LAMP reaction devices (Diagenetix, Inc., Honolulu, HI, USA), connected to Android 4.4 enabled, Bluetooth-capable Nexus 7 tablets for output. Quantification by a quantitative PCR assay was assumed correct to compare the lab and grower qLAMP assay quantification. Growers were able to conduct and interpret qLAMP results; however, theErysiphe necatorinoculum quantification was unreliable using the beta-Smart-DART devices. The qLAMP assay developed was sensitive to one spore in early testing of the assay, but decreased to >20 spores by the end of the trial. The qLAMP assay is not likely a suitable management tool for grape powdery mildew due to losses in sensitivity and decreasing costs and portability for other, more reliable molecular tools.

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Hu, Lianxia, Shufei Zhang, Yuling Xue, Yaoguang Zhang, Wei Zhang et Shijie Wang. « Quantitative Detection of Viable but Nonculturable Cronobacter sakazakii Using Photosensitive Nucleic Acid Dye PMA Combined with Isothermal Amplification LAMP in Raw Milk ». Foods 11, n^o 17 (1 septembre 2022) : 2653. http://dx.doi.org/10.3390/foods11172653.

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An accurate method that rapidly detects the number of viable but nonculturable (VBNC) Cronobacter sakazakii was developed by combining propidium bromide with quantitative LAMP (PMA-QLAMP). The gyrB gene was the target for primers design. The optimal PMA treatment conditions were determined to eliminate the DNA amplification of 108 CFU/mL of dead C. sakazakii without affecting any viable C. sakazakii DNA amplification. Compared with the DNA of 24 strains of common non-C. sakazakii strains found in raw milk and dairy products, the DNA of only six C. sakazakii strains from different sources was amplified using PMA-QLAMP. The ability of PMA-QLAMP to quantitatively detect non-dead C. sakazakii in a 10% powdered infant formula (PIF) solution was limited to 4.3 × 102 CFU/mL and above concentrations. Pasteurizing 106 CFU/mL viable C. sakazakii yielded the maximum ratio of the VBNC C. sakazakii. PMA-QLAMP-based detection indicated that, although approximately 13% of 60 samples were positive for viable C. sakazakii, the C. sakazakii titers in these positive samples were low, and none entered the VBNC state under pasteurization. PMA-QLAMP showed potential as a specific and reliable method for detecting VBNC-C. sakazakii in pasteurized raw milk, thereby providing an early warning system that indicates potential contamination of PIF.

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Lee, Min-Young, Vu-Minh Phan, Woo-In Lee, Yee-Hyung Kim, Sung-Wook Kang et Tae-Seok Seo. « Developing a Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Seven Respiratory Viruses including SARS-CoV-2 ». Medicina 58, n^o 9 (5 septembre 2022) : 1224. http://dx.doi.org/10.3390/medicina58091224.

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Background and Objectives: The coronavirus disease (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), continues to be a pandemic even in 2022. As the initial symptoms of COVID-19 overlap with those of infections from other respiratory viruses, an accurate and rapid diagnosis of COVID-19 is essential for administering appropriate treatment to patients. Currently, the most widely used method for detecting respiratory viruses is based on real-time polymerase chain reaction (PCR) and includes reverse-transcription real-time quantitative PCR (RT-qPCR). However, RT-qPCR assays require sophisticated facilities and are time-consuming. This study aimed to develop a real-time quantitative loop-mediated isothermal amplification (RT-qLAMP) assay and compare its analytical performance with RT-qPCR. Materials and Methods: A total of 315 nasopharyngeal swabs from patients with symptoms of respiratory infections were included in this study. A primary screening of the specimens was performed using RT-qPCR. RNA/DNA from standard strains for respiratory viruses and heat-inactivated preparations of standard strains for SARS-CoV-2 were used to evaluate the accuracy and target specificity of the RT-qLAMP assay. Results: We successfully developed an RT-qLAMP assay for seven respiratory viruses: respiratory syncytial virus (RSV) A, RSV B, adenovirus, influenza (Flu) A (H1N1 and H3N2), Flu B, and SARS-CoV-2. RT-qLAMP was performed in a final reaction volume of 9.6 µL. No cross-reactivity was observed. Compared with the RT-PCR results, the sensitivity and specificity of the RT-qLAMP assay were 95.1% and 100%, respectively. The agreement between the two methods was 97.1%. The median amplification time to RT-qLAMP positivity was 22:34 min (range: 6:80–47:98 min). Conclusions: The RT-qLAMP assay requires a small number of reagents and samples and is performed with an isothermal reaction. This study established a fast, simple, and sensitive test that can be applied to point-of-care testing devices to facilitate the detection of respiratory viruses, including SARS-CoV-2.

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Kumsiri, Ratchanok, et Panan Kanchanaphum. « A Comparison of Four Molecular Methods for Detection of Aflatoxin-Producing Aspergillus in Peanut and Dried Shrimp Samples Collected from Local Markets around Pathum Thani Province, Thailand ». Scientifica 2020 (24 décembre 2020) : 1–6. http://dx.doi.org/10.1155/2020/8580451.

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Aspergillus flavus is an aflatoxin-producing fungus which is poisonous to humans and animals when consumed. Detecting the fungus can help to prevent this danger. The four molecular methods, namely, conventional isothermal amplification (LAMP), PCR, quantitative LAMP (qLAMP), and qPCR, were compared to determine their efficiency for A. flavus detection. Thirty samples of peanut and dried shrimp were collected from 15 markets around Pathum Thani Province in Thailand. The samples were artificially infected with 108 conidia/ml of A. flavus for 1 hr and enriched for one day to represent real contamination. The results show that the sensitivity detection for A. flavus in PCR, LAMP, qPCR, and qLAMP was 50 ng, 5 ng, 5 pg, and 5 pg, respectively. Aspergillus in 30 peanut and dried shrimp from the market was detected by all four methods. The detection rate was about 20%, 60%, 100%, and 100% with PCR, LAMP, qPCR, and qLAMP, respectively. The molecular detection technique, especially LAMP, qPCR, and qLAMP, can detect this pathogenic fungi very rapidly with high sensitivity and reliability in comparison to conventional PCR.

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Vichaibun, Virun, et Panan Kanchanaphum. « Quantitative LAMP and PCR Detection of Salmonella in Chicken Samples Collected from Local Markets around Pathum Thani Province, Thailand ». International Journal of Food Science 2020 (10 juillet 2020) : 1–6. http://dx.doi.org/10.1155/2020/8833173.

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Salmonella is a bacterium that infects people when they consume contaminated food or liquids. To prevent humans from becoming ill, it is useful to have an efficient method of detecting Salmonella before the disease is passed on through the food chain. In this research, the efficiency of Salmonella detection was compared using the following four methods: conventional loop-mediated isothermal amplification (LAMP), PCR, quantitative LAMP (qLAMP), and qPCR. The artificial infection of chicken samples started with incubating of 10 mL of 108 CFU of S. typhimurium for 6 hr. and enriching for 2 hr. to represent real contamination of the samples. The results show that the sensitivity of Salmonella DNA detection in PCR, qPCR, LAMP, and qLAMP were 50 ng, 5 ng, 50 pg, and and 500 fg, respectively. Thirty samples of 10 g chicken were collected from 10 markets in Pathum Thani, Thailand; then, the infection was detected. The conventional LAMP, qLAMP, and qPCR methods detected Salmonella in all the chicken samples. However, the conventional PCR method detected Salmonella infection in only eight of the samples. Overall, the qLAMP method had the highest sensitivity of Salmonella DNA detection.

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Villari, Caterina, Walter F. Mahaffee, Thomas K. Mitchell, Kerry F. Pedley, Michael L. Pieck et Francesca Peduto Hand. « Early Detection of Airborne Inoculum of Magnaporthe oryzae in Turfgrass Fields Using a Quantitative LAMP Assay ». Plant Disease 101, n^o 1 (janvier 2017) : 170–77. http://dx.doi.org/10.1094/pdis-06-16-0834-re.

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Gray leaf spot (GLS) is a destructive disease of perennial ryegrass caused by a host specific pathotype of the ascomycete Magnaporthe oryzae. Early diagnosis is crucial for effective disease management and the implementation of Integrated Pest Management practices. However, a rapid protocol for the detection of low levels of airborne inoculum is still missing. We developed a pathogen-specific quantitative loop-mediated isothermal amplification (qLAMP) assay coupled with a spore trap system for rapid detection and quantification of airborne inoculum of the M. oryzae perennial ryegrass pathotype, and tested its suitability for implementation in GLS-infected turfgrass fields. In summer 2015, two perennial ryegrass plots were artificially inoculated with the pathogen, with four continuously running custom impaction spore traps placed in each plot. Sampling units were replaced daily and tested with the developed qLAMP assay, while plots were monitored for symptom development. Results confirmed that the qLAMP assay-trap system was able to detect as few as 10 conidia up to 12 days before symptoms developed in the field. LAMP technology is particularly appropriate for field implementation by nontechnical users, and has the potential to be a powerful decision support tool to guide timing of fungicide applications for GLS management.

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Grabicoski, Edilaine Mauricia Gelinski, David de Souza Jaccoud-Filho, David Lee, Luciane Henneberg et Marcos Pileggi. « Real-Time Quantitative and Ion-Metal Indicator LAMP-Based Assays for Rapid Detection of Sclerotinia sclerotiorum ». Plant Disease 104, n^o 5 (mai 2020) : 1514–26. http://dx.doi.org/10.1094/pdis-07-19-1455-re.

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Sclerotinia sclerotiorum is one of the most devastating and cosmopolitan plant pathogens. Rapid detection of S. sclerotiorum can provide growers an advantage in knowing what control measures should be taken to minimize crop damage and financial losses caused by it. Loop-mediated isothermal amplification (LAMP) is a fast, sensitive, and specific nucleic acid amplification method that does not require a thermal cycler. This study aimed to develop a LAMP-based assay for the specific detection of S. sclerotiorum (Ss-LAMP). A real-time quantitative LAMP reaction (Ss-qLAMP) and a calcein ion indicator-based LAMP reaction (Ss-cLAMP) were designed, optimized, and tested on fungi, plant, and soil samples. The Ss-LAMP reactions were very specific and sensitive. Applying the artificially inoculated soil samples with DNA purified by five protocols in the Ss-qLAMP reaction, it was possible to detect and quantify the pathogen DNA, regardless of the extraction protocol. Naturally infected soybean tissues had the pathogen detected by Ss-cLAMP directly in the reaction tube with no DNA extraction requirement. The assays should be applicable for many types of samples, such as soil, spore traps, and plant tissues from several crops, with no requirement for DNA extraction. The Ss-LAMP reactions took less than 1 h to complete, and they can be made directly in the field with real-time quantitative results (Ss-qLAMP) or qualitative naked-eye visual results (Ss-cLAMP). Results were obtained with 10 pg of DNA or 10 ng of crude mycelium, suggesting a detection limit close to a single DNA copy. Ss-LAMP reactions will allow rapid and accurate diagnosis of S. sclerotiorum and assist in pathogen management and control.

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Chen, Bo-Jun, Veerappan Mani, Sheng-Tung Huang, Yi-Chiuen Hu et His-Chi Peter Shan. « Bisintercalating DNA redox reporters for real-time electrochemical qLAMP ». Biosensors and Bioelectronics 129 (mars 2019) : 277–83. http://dx.doi.org/10.1016/j.bios.2018.09.056.

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Ongerth, Jerry E., et Richard E. Danielson. « RT qLAMP—Direct Detection of SARS-CoV-2 in Raw Sewage ». Journal of Biomolecular Techniques : JBT 32, n^o 3 (septembre 2021) : 206–13. http://dx.doi.org/10.7171/jbt.21-32-03-016.

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Kumsiri, Ratchanok, et Panan Kanchanaphum. « Comparison of Time Course Detection of Human Male DNA from Blood Stains on Various Objects on Surface in a Natural Environment and in a Laboratory Using Loop-Mediated Isothermal Amplification (LAMP) ». Scientifica 2021 (25 novembre 2021) : 1–7. http://dx.doi.org/10.1155/2021/4811608.

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In forensic study, the biological evidence can easily degrade, especially DNA. Degraded and environmentally challenged samples can produce numerous problems in forensic DNA analysis including loss of band product. Loop-mediated isothermal amplification or LAMP is one of the DNA analysis techniques used in forensic study. This study explores the limitations of the efficiency of the LAMP technique on abandoned DNA. For the DNA template, 8 male and 2 female blood-stained samples were taken from the surfaces, namely, brick, cloth, and tile from inside, and buried outside the laboratory. The LAMP reaction was used to amplify the SRY gene for detecting male DNA. All the blood-stained samples were stored for 1, 7, 15, 30, and 45 day (s). The LAMP product from the blood-stained samples on all the surfaces that were kept in a laboratory was detected using the gel electrophoresis technique from day 1 until day 45. However, the LAMP product on day 30 and 45 was smear and dim. The LAMP product from the blood-stained samples buried outside the laboratory was observed using the gel electrophoresis technique within day 30 (smear and dim). To increase the efficiency of detection, the qLAMP technique detected product on all the male samples from all the surfaces buried outside the laboratory for 45 days. The results indicate that this LAMP condition was possible detecting male DNA and the environmental factors are the main influence on the sensitivity of the LAMP technique. In addition, the qLAMP technique can increase the capacity and sensitivity of the detection.

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Thèses sur le sujet "QLAMP"

PULTRONE, CINZIA. « Sviluppo di una tecnologia Q-LAMP quantitativa mediante la messa a punto di un nuovo modello di saggio per la rilevazione e la quantificazione del virus Epstein-Barr ». Doctoral thesis, Università degli Studi di Milano-Bicocca, 2016. http://hdl.handle.net/10281/138662.

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Epstein-Barr virus (EBV) also called human herpesvirus 4 (HHV-4) is a double-stranded DNA viruses that infects B cells. The genome is about 172 kb in length and encodes for more than 80 genes. EBV is classified into two subtypes: Type 1 / A and 2 / B depending on the sequence of the EBNA-2 gene. More than 90% of the world’s population has been exposed to the EBV infection, like all herpes viruses it is able to persist in the host for life with a silent infection by integrating in its genome. The quantification of EBV DNA loads in blood, plasma, serum and CSF (cerebrospinal fluid) is essential for the diagnosis of chronic infection of EBV, especially for the diagnosis of EBV-associated lymphoproliferative disorders in immunocompromised subjects. Currently serological assay and methods PCR-based has been used for the detection of EBV. Both of these are time consuming and need more steps for the set-up, therefore require specialized personnel and equipped laboratories. This thesis describes the development of a quantitative Q-LAMP technology generating a new model assay for the detection and quantification of the two most clinically relevant subtypes of EBV virus. Initially the Q-LAMP technology has been developed only for qualitative purposes thanks to its features to be simple, rapid, specific and sensitive, therefore it is easily applicable in the clinical laboratories by using basic facilities. The Q-LAMP EBV assay has been designed using the qualitative Q-LAMP properties and has been improved to be quantitative, high sensitive, specific and accurate on four different matrices (whole blood, plasma, serum and CSF). The optimization has been carried out both on plasmids and on negative extracted matrices and the final assay has been validated on clinical samples. Thanks to the multiplex format it has been possible to introduce in the reaction a control that allows the assessment of the sample extraction and the validation of the negative result. The amplification and the detection of the target and the internal control occur in real-time in a single tube. To evaluate the diagnostic performance and the feasibility in clinical practice, the assay has been validated on positive and negative clinical samples compared with the standard method Real-Time PCR using as extraction method the Liaison IXT platform (DiaSorin). The correlation between the two methods on whole blood and plasma is acceptable and meet the requests of corporate guidelines, made in response to the market trend. These results have been demonstrated that Q-LAMP EBV assay is suitable for industrialization for a future diagnostic product.

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Actes de conférences sur le sujet "QLAMP"

Tanusy, Jeanyfer. « Feminism in Roald Dahl's qLamb to the Slaughterq : A Semiotic Analysis ». Dans Fourth Prasasti International Seminar on Linguistics (Prasasti 2018). Paris, France : Atlantis Press, 2018. http://dx.doi.org/10.2991/prasasti-18.2018.30.

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