Detection methods for milk pathogenic bacteria by loop-mediated isothermal amplification

Yang, Wentao; Song, Xiaoning; Wang, Jingxin; Li, Zhen; Ji, Mingjiang; Li, Yufeng

doi:10.5582/bst.2014.01118

Cited by 10 publications

(7 citation statements)

References 20 publications

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“…Furthermore, actual samples from poultry farms were tested directly with the LAMP assays, and in all types of the samples tested, APEC-associated virulence genes were detected. The applicability of this LAMP method to direct detection of E. coli has been reported previously for beef and bovine feces samples (Stratakos et al, 2017), minced meat (Ravan et al, 2016), milk (Yang et al, 2014), lettuce (Xue-han et al, 2013), stool samples (Song et al, 2005; Teh et al, 2014), and others. In the present study, we have shown that this LAMP method is also applicable for direct testing of various environmental samples (e.g., feed, feces, air sampled in PBS buffer) as well as of tissues of the internal organs of chickens.…”

Section: Discussionsupporting

confidence: 53%

Loop-mediated isothermal amplification: rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coliin poultry

et al. 2019

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Infections with pathogenic Escherichia coli can lead to different animal- and human-associated diseases. E. coli infections are common in intensive poultry farming, and important economic losses can be expected during infections with avian pathogenic E. coli (APEC) strains followed by colibacillosis. Loop-mediated isothermal amplification (LAMP) assays were developed for rapid detection of 3 APEC-associated virulence genes: sitA, traT, and ompT. All 3 LAMP assays are shown to be specific, repeatable, and reproducible. High sensitivities of the assays are shown, where as few as 1,000 bacterial cells/mL can be detected in different matrices. On-site applicability of this LAMP method is demonstrated through testing of different sample types, from animal swabs and tissues, and from environmental samples collected from 6 commercial poultry farms. All 3 virulence genes were detected at high rates (above 85%) in samples from layer and broiler chickens with clinical signs and, interestingly, high prevalence of those genes was detected also in samples collected from clinically healthy broiler flock (above 75%) while lower prevalence was observed in remaining 3 clinically healthy chicken flocks (less than 75%). Importantly, these virulence genes were detected in almost all of the air samples from 11 randomly selected poultry houses, indicating air as an important route of E. coli spread. Three LAMP assays that target APEC-associated virulence genes are shown to be sensitive and robust and are therefore applicable for rapid on-site testing of various sample types, from animal swabs to air. This on-site LAMP testing protocol offers rapid diagnostics, with results obtained in <35 min, and it can be applied to other important microorganisms to allow the required prompt measures to be taken.

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Section: Discussionsupporting

confidence: 53%

Loop-mediated isothermal amplification: rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coliin poultry

et al. 2019

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“…A previous report demonstrated that the sensitivity of the LAMP assay for Shiga-toxin-producing E. coli (based on Shiga toxin genes, other virulence genes, and O-antigen gene clusters) was between 1 and 20 cells per reaction in pure culture and 10 3 and 10 4 CFU/g in vegetables 26 . The sensitivity of LAMP for the detection of enterotoxigenic E. coli in raw milk was 547 CFU/ml 27 .…”

Section: Discussionmentioning

confidence: 99%

Rapid and sensitive enumeration of total coliforms and Escherichia coli in water and foods by most-probable-number loop-mediated isothermal amplification (MPN-LAMP) method

Tongphrom¹,

Preeprem²,

Nishibushi³

et al. 2018

ScienceAsia

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Total coliforms and Escherichia coli are bacterial indicators used to assess the microbiological qualities of water and foods. In this study, we developed a most-probable-number loop-mediated isothermal amplification (most probable number (MPN-LAMP)) method for the enumeration of total coliforms and E. coli in drinking water and vegetables. The LAMP primers were designed based on lacZ (lacZ-LAMP) and uidA (uidA-LAMP) genes for the detection of total coliforms and E. coli, respectively. In pure culture, the lacZ-LAMP and the uidA-LAMP were able to detect all of the 37 coliforms and 30 E. coli strains, respectively. In the artificially contaminated drinking water and vegetables, the combination of MPN and LAMP techniques (MPN-LAMP) was able to detect E. coli at 1 colony-forming unit (CFU)/100 ml and 5 CFU/g, respectively. From analysis of 33 drinking water and 46 vegetable samples, the total coliform detection obtained by the MPN-LAMP method was in agreement with the MPN detection technique. However, MPN-LAMP was more sensitive than the MPN technique for E. coli detection. Our findings revealed that the MPN-LAMP assay was more rapid and highly sensitive than MPN method. Thus this method could be considered for detection and enumeration of total coliforms and E. coli in the food industry.

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“…The assay used centrifugation and PMA pretreatment and could detect 6.1 × 10 3 to 6.1 × 10 4 CFU/g in 3 h. A study used LAMP to detect V. parahaemolyticus in shrimp utilizing centrifugation as a pretreatment (Yamazaki and others ). The assay could detect 5.3 × 10 2 CFU/g in 1 h. Another study used a LAMP assay to detect enterotoxigenic E. coli from raw milk (Yang and others ). The assay was capable of detecting 547 CFU/mL of the pathogen.…”

Section: Nucleic Acid‐based Detection Methodsmentioning

confidence: 99%

Culture‐Independent Rapid Detection Methods for Bacterial Pathogens and Toxins in Food Matrices

Wang

Salazar

2015

Comp Rev Food Sci Food Safe

228

152

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Rapid detection of bacterial pathogens and toxins in foods is necessary to provide real-time results to mitigate foodborne illness outbreaks. Cultural enrichment methods, although the most widely used, are time-consuming and therefore inadequate for rapid pathogen detection from food samples. The development of novel "rapid" detection methods has decreased detection time dramatically. This review presents an overview of detection methods for various foodborne pathogens, including Listeria monocytogenes, Salmonella enterica, and shiga toxin-producing Escherichia coli, and bacterial toxins in food matrices, with emphasis on those methods which do not require cultural enrichment. Discussed methods include nucleic acid-, immunological-, and biosensor-based techniques. A summary of each type of detection method is given, including referenced methods from the literature. Since these discussed methods do not require cultural enrichment, there is a higher probability of interference from the food matrices. Therefore, the review also discusses the potential interference of food components on detection methods and addresses preprocessing strategies to overcome matrix associated inhibition and to concentrate low quantities of pathogens and toxins in food. Development of rapid and sensitive detection technologies advances and ensures public health safety and security.

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Detection methods for milk pathogenic bacteria by loop-mediated isothermal amplification

Cited by 10 publications

References 20 publications

Loop-mediated isothermal amplification: rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coliin poultry

Loop-mediated isothermal amplification: rapid molecular detection of virulence genes associated with avian pathogenic Escherichia coliin poultry

Rapid and sensitive enumeration of total coliforms and Escherichia coli in water and foods by most-probable-number loop-mediated isothermal amplification (MPN-LAMP) method

Culture‐Independent Rapid Detection Methods for Bacterial Pathogens and Toxins in Food Matrices

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