Articles de revues : « QLAMP »

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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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Salazar, Andrea, Francisco M. Ochoa-Corona, Jennifer D. Olson, Binoy Babu et Mathews Paret. « Probing Loop-Mediated Isothermal Amplification (LAMP) targeting two gene-fragments of rose rosette virus ». PLOS ONE 16, n^o 11 (29 novembre 2021) : e0256510. http://dx.doi.org/10.1371/journal.pone.0256510.

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This study explores the development of Loop-mediated isothermal amplification (LAMP) for detection of rose rosette virus (RRV), a technique with the potential to be translated to rose nurseries. RRV is a negative-sense, single-stranded RNA virus which is a member of the genus Emaravirus (Family Fimoviridae) and the causal agent of the rose rosette disease (RRD). Although RRV symptoms are characteristics, early visual diagnosis of RRD can be misleading and confusing since it may appear like herbicide damage. Moreover, it may take incubation time for symptoms to appear after virus infection. Two sets of RRV gene sequences RNA3 and RNA4 were analyzed and two sets of four LAMP primers were designed. The direct antigen-capture method for direct trapping of RRV in plastic was used for RNA extraction followed by cDNA synthesis. RT-LAMP reactions were for 1 hour at 64°C (RRV-P3) and 66.5°C (RRV-P4) using either a thermocycler or a portable dry bath. RT-qLAMP was also optimized using DNA polymerase GspSSD LD using the same RRV sets of primers. RRV was detected in symptomatic and non-symptomatic RRD tissue from Oklahoma. The limit of detection (LoD) was 1pg/μL and 1 fg/μL using Bst 2.0 LAMP and GspSSD LD quantitative LAMP, respectively. In visual colorimetric pre- and post-reactions, the LoD was 10 pg/μL and 0.1 pg/μL using hydroxy naphthol blue (HNB, 120 μM) and SYBR green I (1:10 dilution), respectively. No cross-reactivity was detected in the RT-LAMP reaction testing cDNAs of eight commonly co-infecting rose viruses and one virus taxonomically related to RRV. Four different dyes were tested, and visible colorimetric reactions were obtained with RT-LAMP Bst 2.0 combined with SYBR I or HNB. RT-qLAMP with GspSSD2.0 offers LoD equal to RT-PCR and it is faster since it works with RNA directly.

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Gong, Han-yue, Qing-yang Li, Huang Zhang, Lei Ye, Lei Shi et Yong-hui Feng. « Development and comparison of qPCR and qLAMP for rapid detection of the decapod iridescent virus 1 (DIV1) ». Journal of Invertebrate Pathology 182 (juin 2021) : 107567. http://dx.doi.org/10.1016/j.jip.2021.107567.

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Dou, Maowei, Sharma T. Sanjay, Delfina C. Dominguez, Sihui Zhan et XiuJun Li. « A paper/polymer hybrid CD-like microfluidic SpinChip integrated with DNA-functionalized graphene oxide nanosensors for multiplex qLAMP detection ». Chemical Communications 53, n^o 79 (2017) : 10886–89. http://dx.doi.org/10.1039/c7cc03246c.

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Telli, A. Ezgi, et Yusuf Doğruer. « Discrimination of viable and dead Vibrio parahaemolyticus subjected to low temperatures using Propidium Monoazide – Quantitative loop mediated isothermal amplification (PMA-qLAMP) and PMA-qPCR ». Microbial Pathogenesis 132 (juillet 2019) : 109–16. http://dx.doi.org/10.1016/j.micpath.2019.04.029.

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Garrido-Maestu, Alejandro, Sarah Azinheiro, Pablo Fuciños, Joana Carvalho et Marta Prado. « Highly sensitive detection of gluten-containing cereals in food samples by real-time Loop-mediated isothermal AMPlification (qLAMP) and real-time polymerase chain reaction (qPCR) ». Food Chemistry 246 (avril 2018) : 156–63. http://dx.doi.org/10.1016/j.foodchem.2017.11.005.

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Alvarez, Mario Moisés, Sergio Bravo-González, Everardo González-González et Grissel Trujillo-de Santiago. « Portable and Label-Free Quantitative Loop-Mediated Isothermal Amplification (LF-qLamp) for Reliable COVID-19 Diagnostics in Three Minutes of Reaction Time : Arduino-Based Detection System Assisted by a pH Microelectrode ». Biosensors 11, n^o 10 (13 octobre 2021) : 386. http://dx.doi.org/10.3390/bios11100386.

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Loop-mediated isothermal amplification (LAMP) has been recently studied as an alternative method for cost-effective diagnostics in the context of the current COVID-19 pandemic. Recent reports document that LAMP-based diagnostic methods have a comparable sensitivity and specificity to that of RT-qPCR. We report the use of a portable Arduino-based LAMP-based amplification system assisted by pH microelectrodes for the accurate and reliable diagnosis of SARS-CoV-2 during the first 3 min of the amplification reaction. We show that this simple system enables a straightforward discrimination between samples containing or not containing artificial SARS-CoV-2 genetic material in the range of 10 to 10,000 copies per 50 µL of reaction mix. We also spiked saliva samples with SARS-CoV-2 synthetic material and corroborated that the LAMP reaction can be successfully monitored in real time using microelectrodes in saliva samples as well. These results may have profound implications for the design of real-time and portable quantitative systems for the reliable detection of viral pathogens including SARS-CoV-2.

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Caqueo-Urízar, Alejandra, Marine Alessandrini et Laurent Boyer. « Calidad de vida en pacientes con esquizofrenia de origen Aymara en la zona Centro-Sur de los Andes ». Universitas Psychologica 16, n^o 5 (29 janvier 2018) : 1–13. http://dx.doi.org/10.11144/javeriana.upsy16-5.qlap.

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El objetivo de este estudio consistió en comparar la calidad de vida (CV) de pacientes con esquizofrenia pertenecientes al grupo étnico aymara de los Andes Centro-Sur, con pacientes no Aymara. En este estudio transversal participaron 253 pacientes de tres clínicas de salud mental en Chile, Perú y Bolivia. Se recogieron datos sociodemográficos y características clínicas. La calidad de vida se evaluó utilizando el Cuestionario S-QoL18. Los análisis comparativos exploraron las diferencias de QoL entre los pacientes Aymara y no Aymara. Los participantes de origen Aymara tuvieron puntuaciones de CV más bajos en comparación con los pacientes no Aymara para el Índice total, las relaciones familiares y la dimensión de vida sentimental. Los ingresos familiares mensuales y la duración del trastorno fueron significativamente más bajos en los pacientes Aymara. Nuestro estudio soporta la hipótesis de una peor CV en pacientes aymaras con esquizofrenia.

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Cao, Zhi, Ke Zhang, Dehua Yin, Qiaoya Zhang, Ying Yu, Jianxin Wen et Hongbo Ni. « Clinical validation of visual LAMP and qLAMP assays for the rapid detection of Toxoplasma gondii ». Frontiers in Cellular and Infection Microbiology 12 (26 septembre 2022). http://dx.doi.org/10.3389/fcimb.2022.1024690.

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Humans are exposed to Toxoplasma gondii infection as pet cats gradually become family members and represent an increasing public health risk worldwide. Toxoplasmosis diagnosis constitutes an important measure for disease prevention and control. In this study, real-time fluorescence quantitative loop-mediated isothermal amplification (qLAMP) and visual LAMP detection technologies were established to conduct tests of T. gondii based on the membrane DNA extraction method, and the optimal detection mix was determined by adding the protective reagent trehalose and screening the concentrations of Mg2+ and dNTPs. Paraffin and lyophilization were used to reduce and even remove aerosol pollution, constructing a detailed anti-contamination protocol. Based on the positive standard plasmid DNA, the LODs of qLAMP and visual LAMP were 92 copies/μL and 92 copies/μL, and the standard curve of qLAMP was Y=2.9503X+20.8992 with R2 = 0.99. The applicability of the qLAMP and visual LAMP assays in disease diagnosis was assessed by evaluating 200 clinical cat faeces samples. The assays showed good diagnostic consistency, with kappa values of 1.0 and 0.99 compared with TaqMan qPCR, respectively. Compared with TaqMan qPCR, the diagnostic specificity/sensitivity of qLAMP and visual LAMP were 100%/100% and 100%/80%, respectively. The qLAMP and visual LAMP assays reported here are rapid and simple tests without extensive sample preparation and have a short turnaround time within 60 min, making them suitable for point-of-care testing.

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Hu, Lianxia, Yuling Xue, liru Cui, Dong Zhang, Lili Feng, Wei Zhang et Shijie Wang. « Detection of viable Lacticaseibacillus paracasei in fermented milk using propidium monoazide combined with quantitative loop-mediated isothermal amplification ». FEMS Microbiology Letters 368, n^o 20 (octobre 2021). http://dx.doi.org/10.1093/femsle/fnab148.

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ABSTRACT To quantify viable probiotic Lacticaseibacillus paracasei (L. paracasei) in fermented milk accurately and quickly, propidium monoazide combined with quantitative loop-mediated isothermal amplification (PMA-qLAMP) was applied. The optimal PMA treatment conditions for treating a L. paracasei suspension were determined using an orthogonal test to eliminate the DNA amplification of 108 CFU/mL of dead L. paracasei. Primers were designed based on the species-specific gyrB gene of L. paracasei. A phylogenetic tree based on the gyrB gene showed that L. paracasei clustered on the same branch with 91% support. Compared with the 16 strains commonly found in fermented milk, three strains of L. paracasei showed positive PMA-qLAMP results, and the melting temperature was approximately 82.4°C. There was a linear relationship (R2 = 0.9983) between the Ct values and the logarithm of the concentration of viable bacteria. The PMA-qLAMP detection limit for the L. paracasei artificially added to fermented milk was 7.3 × 102 CFU/mL. There was no significant difference between the logarithm values of the concentration of viable L. paracasei of 50 fermented milk samples within shelf life using the PMA-qLAMP and plate count methods (P > 0.01). PMA-qLAMP is specific and accurate for obtaining reliable results faster than when using plate counts.

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Zhang, Jingfeng, Li Wang, Lei Shi, Xun Chen, Meidan Liang et Lichao Zhao. « Development and application of a real-time loop-mediated isothermal amplification method for quantification of Acetobacter aceti in red wine ». FEMS Microbiology Letters 367, n^o 19 (octobre 2020). http://dx.doi.org/10.1093/femsle/fnaa152.

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ABSTRACT This study reports the development and optimization of a real-time loop-mediated isothermal amplification (qLAMP) method for rapid detection of Acetobacter aceti strain in red wine samples. Our results showed that the primers and probes designed for 16S rRNA were effective for A. aceti detection. The quantification limit of real-time polymerase chain reaction (qPCR) and qLAMP in pure culture was 2.05 × 101 colony forming units (CFU) mL−1. qLAMP had a sensitivity of 6.88 × 101 CFU mL−1 in artificially contaminated Changyu dry red wine (CDRW) and Changyu red wine (CRW), and 6.88 × 102 CFU mL−1 in artificially contaminated Greatwall dry red wine (GDRW), which was 10 times higher than that of qPCR. In conclusion, this newly developed qLAMP is a reliable, rapid and accurate method for the detection and quantification of A. aceti species in red wine samples. Furthermore, our work provides a standard reference method for the quantitative detection of A. aceti and other acetic acid bacteria during the fermentation and storage of red wine samples.

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Carvalho, Joana, Alejandro Garrido-Maestu, Sarah Azinheiro, Pablo Fuciños, Jorge Barros-Velázquez, Ramón J. De Miguel, Verónica Gros et Marta Prado. « Faster monitoring of the invasive alien species (IAS) Dreissena polymorpha in river basins through isothermal amplification ». Scientific Reports 11, n^o 1 (13 mai 2021). http://dx.doi.org/10.1038/s41598-021-89574-w.

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AbstractZebra mussel (Dreissena polymorpha) is considered as one of the 100 most harmful IAS in the world. Traditional detection methods have limitations, and PCR based environmental DNA detection has provided interesting results for early warning. However, in the last years, the development of isothermal amplification methods has received increasing attention. Among them, loop-mediated isothermal amplification (LAMP) has several advantages, including its higher tolerance to the presence of inhibitors and the possibility of naked-eye detection, which enables and simplifies its potential use in decentralized settings. In the current study, a real-time LAMP (qLAMP) method for the detection of Dreissena polymorpha was developed and tested with samples from the Guadalquivir River basin, together with two real-time PCR (qPCR) methods using different detection chemistries, targeting a specific region of the mitochondrial gene cytochrome C oxidase subunit I. All three developed approaches were evaluated regarding specificity, sensitivity and time required for detection. Regarding sensitivity, both qPCR approaches were more sensitive than qLAMP by one order of magnitude, however the qLAMP method proved to be as specific and much faster being performed in just 9 min versus 23 and 29 min for the qPCR methods based on hydrolysis probe and intercalating dye respectively.

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Oscorbin, Igor P., Ekaterina A. Belousova, Aleksandr I. Zakabunin, Ulyana A. Boyarskikh et Maksim L. Filipenko. « Comparison of fluorescent intercalating dyes for quantitative loop-mediated isothermal amplification (qLAMP) ». BioTechniques 61, n^o 1 (1 juillet 2016). http://dx.doi.org/10.2144/000114432.

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Duarte-Guevara, Paula, Carlos Duarte-Guevara, Akid Ornob et Rashid Bashir. « On-chip PMA labeling of foodborne pathogenic bacteria for viable qPCR and qLAMP detection ». Microfluidics and Nanofluidics 20, n^o 8 (19 juillet 2016). http://dx.doi.org/10.1007/s10404-016-1778-2.

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Yu, Yejiong, Johnny X. Y. Zhou, Binbin Li, Mengmeng Ji, Yun Wang, Emma Carnaby, Monique I. Andersson, Wei E. Huang et Zhanfeng Cui. « A quantitative RT‐qLAMP for the detection of SARS‐CoV ‐2 and human gene in clinical application ». Microbial Biotechnology, 13 juillet 2022. http://dx.doi.org/10.1111/1751-7915.14112.

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Soares-Santos, Verónica, Isabel Pardo et Sergi Ferrer. « Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP ». Frontiers in Microbiology 9 (17 août 2018). http://dx.doi.org/10.3389/fmicb.2018.01945.

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DOĞRUER, Yusuf, et A. Ezgi Telli. « Determination of Vibrio parahaemolyticus in seafoods using direct plate counting, quantitative loop-mediated isothermal amplification and propidium monoazide-qLAMP ». Ankara Üniversitesi Veteriner Fakültesi Dergisi, 1 septembre 2020. http://dx.doi.org/10.33988/auvfd.603868.

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Long, Pan, Zijuan Jiang, Zhengmi He et Zuohong Chen. « Development of a loop-mediated isothermal amplification assay for the rapid detection of Russula subnigricans and Russula japonica ». Frontiers in Microbiology 13 (23 août 2022). http://dx.doi.org/10.3389/fmicb.2022.918651.

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Russula subnigricans is the only deadly species in the genus Russula with a mortality rate of more than 50%, and Russula japonica is the most common poisonous species, making rapid species identification in mushroom poisoning incidents extremely important. The main objective of this study was to develop a rapid, specific, sensitive, and simple loop-mediated isothermal amplification (LAMP) assay for the detection of R. subnigricans and R. japonica. Two sets of species-specific LAMP primers targeting internal transcribed spacer (ITS) regions were designed to identify R. subnigricans and R. japonica. The results demonstrated that while LAMP could specifically detect R. subnigricans and R. japonica, the polymerase chain reaction (PCR) could not distinguish R. subnigricans from Russula nigricans. In addition, the results demonstrated that, compared to electrophoresis-LAMP and real-time quantitative LAMP (RT-qLAMP), the detection sensitivity of HNB-LAMP (a mixture of LAMP with hydroxy naphthol blue (HNB) dye) for R. subnigricans could reach 0.5 pg/μl and was 100-fold higher than that of PCR. The LAMP reaction could be completed in 45 min, which is much faster than the conventional PCR. In the future, LAMP can be used a quick, specific, and sensitive detection tool in various fields.

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Moser, Nicolas, Ling-Shan Yu, Jesus Rodriguez Manzano, Kenny Malpartida-Cardenas, Anselm Au, Paul Arkell, Chiara Cicatiello et al. « Quantitative detection of dengue serotypes using a smartphone-connected handheld lab-on-chip platform ». Frontiers in Bioengineering and Biotechnology 10 (15 septembre 2022). http://dx.doi.org/10.3389/fbioe.2022.892853.

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Dengue is one of the most prevalent infectious diseases in the world. Rapid, accurate and scalable diagnostics are key to patient management and epidemiological surveillance of the dengue virus (DENV), however current technologies do not match required clinical sensitivity and specificity or rely on large laboratory equipment. In this work, we report the translation of our smartphone-connected handheld Lab-on-Chip (LoC) platform for the quantitative detection of two dengue serotypes. At its core, the approach relies on the combination of Complementary Metal-Oxide-Semiconductor (CMOS) microchip technology to integrate an array of 78 × 56 potentiometric sensors, and a label-free reverse-transcriptase loop mediated isothermal amplification (RT-LAMP) assay. The platform communicates to a smartphone app which synchronises results in real time with a secure cloud server hosted by Amazon Web Services (AWS) for epidemiological surveillance. The assay on our LoC platform (RT-eLAMP) was shown to match performance on a gold-standard fluorescence-based real-time instrument (RT-qLAMP) with synthetic DENV-1 and DENV-2 RNA and extracted RNA from 9 DENV-2 clinical isolates, achieving quantitative detection in under 15 min. To validate the portability of the platform and the geo-tagging capabilities, we led our study in the laboratories at Imperial College London, UK, and Kaohsiung Medical Hospital, Taiwan. This approach carries high potential for application in low resource settings at the point of care (PoC).

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Yu, Ning, Shi-Kai Zhang, Jian Chen, Cheng Zhao, Ye-Min Cao, Ling Li et Yong-Bing Cao. « Mitigation of QingLuoTongMai Pills on Chemotherapy-induced Phlebitis : A Network Pharmacology Study and Experimental Validation ». Combinatorial Chemistry & ; High Throughput Screening 25 (29 juin 2022). http://dx.doi.org/10.2174/1386207325666220629121318.

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Objective: Chemotherapy induced phlebitis (CIP) is a side product of chemotherapy treatment for malignant tumors, which affects the therapeutic effect and quality of life of cancer patients, and still lacks a clear therapeutic means. In this study, we investigated the therapeutic effects of QLTMP on CIP using network pharmacology and verified the anti-inflammatory mechanism of QLTMP in mice model induced by vinorelbine. Methods: Network pharmacology analysis was performed to identify bioactive compounds in QLTMP. The protein-protein interaction network was used to identify the core therapeutic targets of QLTMP against CIP. Analyzed biological function and pathway enrichment based on the identified core therapeutic targets. Evaluate the therapeutic effect of QLTMP in a model of CIP induced by vinorelbine to confirm the reliability of the network pharmacological analysis. Materials and methods: The 165 bioactive compounds of QLTMP matched the screening criteria and identified 19 core therapeutic targets of QLTMP against CIP. Biofunctional analysis showed that the therapeutic effect of QLTMP on CIP was mainly related to the inhibition of inflammation; while pathway enrichment analysis showed that TNF signaling pathway was involved in the inflammatory process. Experimental confirmation in mice model showed that QLTMP exerts anti-inflammatory effects through modulation of PI3K/AKT/TNF signaling pathway, a discovery consistent with the network pharmacological analysis. Discussion and conclusions: The network pharmacological analysis of the anti-inflammatory mechanism of QLTMP on CIP and its exploration of in vivo experiments provide a theoretical basis for the design of agents that can mitigate or cure CIP.

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