Rapid Detection of <i>Listeria monocytogenes</i> in Raw Milk with Loop‐Mediated Isothermal Amplification and Chemosensor

Wang, Deguo; Zhang, Gaiping; Chen, Lu; Deng, Ruiguang; AiMin, Zhi; Guo, Junqing; Zhao, Dong; Xu, Zhihong

doi:10.1111/j.1750-3841.2011.02383.x

Cited by 23 publications

(17 citation statements)

References 20 publications

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“…The LAMP reaction can be accelerated by the addition of two loop primers, named loop forward (LF) primer and loop backward (LB) primer (Nagamine et al 2002). By now, loop primers have been successfully applied in many studies (Tomlinson et al 2010a;Wang et al 2011;Moradi et al 2012). However, with the loop primers, the LAMP assay failed to distinguish the resistant mutants from the wild type of F. graminearum in a previous study (Duan et al 2014a).…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of benzimidazole resistance in Botrytis cinerea by loop-mediated isothermal amplification

Fan

Hahn

et al. 2019

Phytopathol Res

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Benzimidazole fungicides (MBCs) have been widely used in agriculture since 1970s, and resistance to this class of fungicides in Botrytis cinerea is reported worldwide. Resistance to MBCs in B. cinerea is related to mutations in the target β-tubulin gene (TUB2). Compared with the mutation at codon 200, the substitutions from glutamic acid to alanine (E198A), valine (E198V), or lysine (E198K) at codon 198 are currently the predominant mutations in MBC resistant populations of B. cinerea. In this study, a loop-mediated isothermal amplification (LAMP) method was established for rapid detection of benzimidazole resistance in B. cinerea. On the basis of the three mutations at TUB2 codon 198, three sets of LAMP primers were designed, and each of these primer sets was able to specifically amplify the DNA containing its corresponding mutation, while no amplification was detected with other mutated or the wild type DNA. After optimization, the sensitivity and specificity tests illustrated that this LAMP assay had good sensitivity and specificity to detect specific resistant genotypes in B. cinerea. Result also showed that the LAMP assay had good reproducibility. Above all, boiled mycelia could be used as templates, which simplified the process and increased the efficiency of the assay. Considering its rapidity, simplicity, and high efficiency, the LAMP assay developed in this study is a promising tool for the diagnosis of benzimidazole resistance in B. cinerea, and will contribute to the monitoring of resistance development to MBCs in the future.

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Section: Discussionmentioning

confidence: 99%

Rapid detection of benzimidazole resistance in Botrytis cinerea by loop-mediated isothermal amplification

Fan

Hahn

et al. 2019

Phytopathol Res

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“…The traditional culture‐based methods remain the ‘gold standard’ for L. monocytogenes detection against which other methods are available, while they are time‐consuming and labor‐intensive, making it unacceptable for rapidly detecting causative pathogens associated with sporadic and outbreaks cases (Aznar & Alarcon, ; Silbernagel et al ., ). A number of PCR, real‐time PCR, and LAMP methods were reported for species‐specific detection of L. monocytogenes , and the primer sequences are mainly based on two gene sequences: hlyA or iap (Liu et al ., ; Tang et al ., ; Wang et al ., , ). However, hlyA and iap are not unique to L. monocytogenes , hlyA is present in L. monocytogenes , L. seeligeri and L. ivanovii , and the iap gene for all six Listeria species (Bubert et al ., ; Gouin et al ., ).…”

Section: Discussionmentioning

confidence: 99%

Rapid and sensitive detection ofListeria monocytogenesby cross-priming amplification oflmo0733gene

Wang

et al. 2014

FEMS Microbiol Lett

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Listeria monocytogenes is a food-borne pathogen that causes severe opportunistic infection in humans and animals. This study reports the development of single cross-priming amplification (S-CPA) and double CPA (D-CPA) assays targeting species-specific gene lmo0733 for identifying L. monocytogenes strains. The CPA assays were performed at a constant temperature 64 °C using seven specific primers and evaluated for specificity and sensitivity. The color change of positive amplification was directly observed by Loopamp Fluorescent Detection Reagent (FD), and the DNA products were visualized as a ladder-like banding pattern on 2.5% gel electrophoresis. Moreover, the positive reactions were also detected by real-time measurement of turbidity. 50 L. monocytogenes and 46 non-L. monocytogenes strains were used for the method verification, and the specificity was 100%. The limit of detection (LoD) of the S-CPA and D-CPA assays was 2.5 pg DNA per reaction and 10-fold more sensitive than PCR. A total of 60 pork samples were tested for L. monocytogenes using the S-CPA assay developed in the study, and the accuracy of the S-CPA and the culture-biotechnical method was 100% identical. The results suggested that the S-CPA assay was a rapid, sensitive, and valuable tool for detection of L. monocytogenes in food products.

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“…Chest CT and RT-PCR has been used for the clinical diagnosis of the coronavirus pneumonia 2, 3 , however, chest CT needs CT equipment and put the professional operator at risk of infection, and RT-PCR has high professional and technical requirements for operators. Loop-mediated isothermal amplification (LAMP) can amplify nucleic acids under isothermal conditions 4 , and it had the advantages over real-time PCR assays in specificity sensitivity, cost effectiveness and rapidity 5,6,7 . The objective of this study was to develop the one-pot real-time RT-LAMP assay and the one-pot visual RT-LAMP assay for diagnosis of the pneumonia caused by COVID-19, which would be suitable for use at less developed areas.…”

Section: Introductionmentioning

confidence: 99%

One-pot Detection of COVID-19 with Real-time Reverse-transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assay and Visual RT-LAMP Assay

Wang

2020

Preprint

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Background: Rapid and reliable diagnostic assays were critical for prevention and control of the coronavirus pneumonia caused by COVID-19. Objective: This study was to establish one-pot real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay and one-pot visual RT-LAMP assay for the detection of COVID-19. Methods: Six specific LAMP primers targeting the N gene of COVID-19 were designed, the RT-LAMP reaction system was optimized with plasmid pUC57 containing N gene sequence, the detection limit was determined with a serial dilution of the plasmid pUC57 containing N gene sequence, and the one-pot real-time RT-LAMP assay and one-pot visual RT-LAMP assay for the detection of COVID-19 were established. Results: Our results showed that the one-pot RT-LAMP assays can detect COVID-19 with a limit of ≥ 6 copies per μl −1 of pUC57 containing N gene sequence. Conclusion: This study provides rapid, reliable and sensitive tools for facilitating preliminary and cost-effective prevention and control of COVID-19.

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Rapid Detection of Listeria monocytogenes in Raw Milk with Loop‐Mediated Isothermal Amplification and Chemosensor

Abstract: The LAMP-chemosensor method reported here provided a powerful tool for detection of L. monocytogenes in raw milk samples due to its specificity, sensitivity, and rapidity.

Cited by 23 publications

References 20 publications

Rapid detection of benzimidazole resistance in Botrytis cinerea by loop-mediated isothermal amplification

Rapid detection of benzimidazole resistance in Botrytis cinerea by loop-mediated isothermal amplification

Rapid and sensitive detection ofListeria monocytogenesby cross-priming amplification oflmo0733gene

One-pot Detection of COVID-19 with Real-time Reverse-transcription Loop-mediated Isothermal Amplification (RT-LAMP) Assay and Visual RT-LAMP Assay

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