Dataset Information

In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats.

ABSTRACT: The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100%, while exclusivity for non-target bacteria was 100%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100%. The RLOD values of the developed mPCR for STEC, L. monocytogenes and Salmonella spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.

SUBMITTER: Jaroenporn C

PROVIDER: S-EPMC9180326 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats.

Jaroenporn Chanokchon C Supawasit Wannakarn W Bundidamorn Damkerng D Udompijitkul Pathima P Assawamakin Anunchai A Trevanich Sudsai S

Foods (Basel, Switzerland) 20220525 11

The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of de ...[more]

PMID: 35681308

Similar Datasets

Project description:The prevalence and levels of Listeria monocytogenes in retail raw foods covering most provincial capitals in China were studied with testing of 1036 samples of vegetables, edible mushrooms, raw meat, aquatic products and quick-frozen products from September 2012 to January 2014. The total prevalence of Listeria monocytogenes was 20.0% (207/1036), and the most probable number (MPN) values of 65.7% of the positive samples ranged from 0.3 to 110 MPN/g. Geographical differences were observed in this survey, and the results of both qualitative and quantitative methods indicated that the levels in the samples from North China were higher than those in the samples from South China. A total of 248 isolates were analyzed, of which approximately half belonged to molecular serogroup 1/2a-3a (45.2%), followed by 1/2b-3b-7 (30.6%), 1/2c-3c (16.1%), 4b-4d-4e (5.2%) and 4a-4c (2.8%). Most of the isolates carried hly (100%), inlB (98.8%), inlA (99.6%), inlC (98.0%) and inlJ (99.2%), and 44.8% of the isolates were llsX-positive. Seventeen epidemic clones (ECs) were detected, with 7 strains belonging to ECI (2.8%) and 10 belonging to ECIII (4.03%). Resistance to clindamycin (46.8%) was commonly observed, and 59 strains (23.8%) were susceptible to all 14 tested antibiotics, whereas 84 (33.9%) showed an intermediate level of resistance or were resistant to two or more antibiotics, including 7 multi-resistant strains that exhibited resistance to more than 10 antibiotics. The data obtained in the present study provides useful information for assessment of the possible risk posed to Chinese consumers, and this information will have a significant public health impact in China. Furthermore, the presence of virulence markers, epidemic clones, as well as the antibiotic resistance amongst the isolates strongly implies that many of these strains might be capable of causing listeriosis, and more accurate treatment of human listeriosis with effective antibiotics should be considered. This research represents a more full-scale and systematical investigation of the prevalence of L. monocytogenes in retail raw foods in China, and it provides baseline information for Chinese regulatory authorities that will aid in the formulation of a regulatory framework for controlling L. monocytogenes with the aim of improving the microbiological safety of raw foods.

Project description:AbstractA comprehensive understanding of foodborne pathogen diversity in preharvest environments is necessary to effectively track pathogens on farms and identify sources of produce contamination. As such, this study aimed to characterize Listeria diversity in wildlife feces and agricultural water collected from a New York state produce farm over a growing season. Water samples were collected from a pond (n = 80) and a stream (n = 52). Fecal samples (n = 77) were opportunistically collected from areas <5 m from the water sources; all samples were collected from a <0.5-km2 area. Overall, 86 (41%) and 50 (24%) of 209 samples were positive for Listeria monocytogenes and Listeria spp. (excluding L. monocytogenes), respectively. For each positive sample, one L. monocytogenes or Listeria spp. isolate was speciated by sequencing the sigB gene, thereby allowing for additional characterization based on the sigB allelic type. The 86 L. monocytogenes and 50 Listeria spp. isolates represented 8 and 23 different allelic types, respectively. A subset of L. monocytogenes isolates (n = 44) from pond water and pond-adjacent feces (representing an ∼5,000-m2 area) were further characterized by pulsed-field gel electrophoresis (PFGE); these 44 isolates represented 22 PFGE types, which is indicative of considerable diversity at a small spatial scale. Ten PFGE types were isolated more than once, suggesting persistence or reintroduction of PFGE types in this area. Given the small spatial scale, the prevalence of L. monocytogenes and Listeria spp., as well as the considerable diversity among isolates, suggests traceback investigations may be challenging. For example, traceback of finished product or processing facility contamination with specific subtypes to preharvest sources may require collection of large sample sets and characterization of a considerable number of isolates. Our data also support the adage "absence of evidence does not equal evidence of absence" as applies to L. monocytogenes traceback efforts at the preharvest level.Highlights

Dataset Information

In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats.

Publications

In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing <i>Escherichia coli</i>, <i>Listeria monocytogenes</i> and <i>Salmonella</i> spp. in Raw Meats.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets