Salmonella in Foods: New Enrichment Procedure for TECRA Salmonella Visual Immunoassay Using a Single RV(R10) Only, TT Only, or Dual RV(R10) and TT Selective Enrichment Broths (AOAC Official Method 998.09): Collaborative Study

Hughes, Denise; Dailianis, Angela; Hill, Louise; Curiale, Michael S; Gangar, Vidhya; Arnold, Douglas R.; Barrat, C; Baxter, Tony; Bell, J B; Brooks, Richard; Bryant, Douglas W.; Burke, Katie L.; Burnie, A. G.; Cliffard, D; Danisavich, T; Daniels, Kourtney A; Deiss, K; Armando, D’Onorio; Faucher, Karla; Finkenbiner, D; Gasanov, Uta; Gebler, J; Gerry, A Quinn; Graham, D. C.; Graham, Todd; Harris, P P; Hetrick, S; Jurgens, Julian F.; Keating, Kathy Jost; Klokman, Renee; Le, C; Matrozza, Mark; McCawley, Cameron; Munyard, S; Pye, V.E.; Rajkowski, Kathleen T.; Ristov, K; Rosinko, Joyce; Schneider, Kathrin; Schubert, Mario; Sloan, Edna M.; Souter, Vivienne L.; Wilson, Maurice; Zuroski, K

doi:10.1093/jaoac/86.4.775

Cited by 7 publications

(4 citation statements)

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“…Although this study tested a limited number of retail meat and poultry samples, it was apparent that RV medium was the superior selective enrichment broth for BAM compared with TT. An earlier study (Hughes et al, 2003 ) did a pairwise comparison of RVR10 and TT for the TECRA Salmonella Visual Immunoassay and did not find a significant difference ( p > 0.05). A recent study (Broadway et al, 2021 ) reported a 1.4% Salmonella prevalence rate (out of 865 retail samples) using the USDA protocol (USDA, 2021), much more effective compared with BAM.…”

Section: Discussionmentioning

confidence: 90%

Toward the Adoption of Loop-Mediated Isothermal Amplification for Salmonella Screening at the National Antimicrobial Resistance Monitoring System's Retail Meat Sites

Young

Domesle

McDonald

et al. 2022

Foodborne Pathogens and Disease

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The National Antimicrobial Resistance Monitoring System (NARMS) is a One Health program in the United States that collects data on antimicrobial resistance in enteric bacteria from humans, animals, and the environment. Salmonella is a major pathogen tracked by the NARMS retail meat arm but currently lacks a uniform screening method. We evaluated a loop-mediated isothermal amplification (LAMP) assay for the rapid screening of Salmonella from 69 NARMS retail meat and poultry samples. All samples were processed side by side for culture isolation using two protocols, one from NARMS and the other one described in the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM). Overall, 10 (14.5%) samples screened positive by the Salmonella LAMP assay. Of those, six were culture-confirmed by the NARMS protocol and six by the BAM method with overlap on four samples. No Salmonella isolates were recovered from samples that screened negative with LAMP. These results suggested 100% sensitivity for LAMP in reference to culture. Antimicrobial susceptibility testing and whole-genome sequencing analysis confirmed identities of these isolates. Using the BAM protocol, all Salmonella isolates were recovered from samples undergoing Rappaport-Vassiliadis medium selective enrichment and presumptive colonies ( n = 130) were dominated by Hafnia alvei (44.6%), Proteus mirabilis (22.3%), and Morganella morganii (9.9%) based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This method comparison study clearly demonstrated the benefit of a rapid, robust, and highly sensitive molecular screening method in streamlining the laboratory workflow. Fourteen NARMS retail meat sites further verified the performance of this assay using a portion of their routine samples, reporting an overall specificity of 98.8% and sensitivity of 90%. As of July 2022, the vast majority of NARMS retail meat sites have adopted the Salmonella LAMP assay for rapid screening of Salmonella in all samples.

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Section: Discussionmentioning

confidence: 90%

Toward the Adoption of Loop-Mediated Isothermal Amplification for Salmonella Screening at the National Antimicrobial Resistance Monitoring System's Retail Meat Sites

Young

Domesle

McDonald

et al. 2022

Foodborne Pathogens and Disease

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“…The next goal was to shorten the screening workflow. As the M broth is considered a post‐enrichment step (Hughes et al ), the real‐time assay was evaluated at the selective enrichment step: the Rappaport‐Vassiliadis R10 (RVR) broth. The results of testing 191 meat samples in RVR broth using the real‐time PCR assay showed 100% concordance when compared to the results of the 3M TECRA kit for the same set of food samples (Table ), similar results were obtained using veterinary samples (Goodman et al ).…”

Section: Resultsmentioning

confidence: 99%

Detection ofSalmonellaspp. in retail meat products: a comparison between a discontinued commercial kit and a laboratory‐developed screening method

Shushe

Wroblewski

MacGowan

et al. 2019

Lett Appl Microbiol

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The National Antimicrobial Resistance Monitoring System for Enteric Bacteria retail food surveillance programme screens retail meat samples for the presence of Salmonella spp. to track antimicrobial resistance in food. In this study, a laboratory developed real‐time PCR assay that detects Salmonella spp. was evaluated as a screening method to replace the discountinued 3M TECRA kit. The 3M TECRA kit was a commercially available, visual immunoassay used to screen food samples for the presence of Salmonella spp. This kit was discontinued in September 2016 by the manufacturer and an alternative screening method was needed to replace the discontinued TECRA kit. Salmonella spp. is detected by the real‐time PCR assay earlier in the screening process than by the TECRA kit. Salmonella spp. can also be reliably isolated from the enrichment broth earlier in the protocol. Additionally, cost analysis shows that the real‐time PCR assay saves $2·50 per sample. New York State Department of Health currently uses this real‐time PCR assay as a screening method for the presence of Salmonella spp. in retail meat samples. The assay allows for continued monitoring of antimicrobial resistance in Salmonella spp., while providing a cost savings and a decrease in turnaround time. Significance and Impact of the Study The National Antimicrobial Resistance Monitoring System for Enteric Bacteria (NARMS) tracks antimicrobial susceptibility of enteric bacteria in people, food and animals (https://www.fda.gov/animalveterinary/safetyhealth/antimicrobialresistance/nationalantimicrobialresistancemonitoringsystem/). The New York State Department of Health (NYSDOH) became a NARMS retail food surveillance (RFS) site in 2003. The NARMS‐RFS programme screens retail meat samples from grocery stores in the United States for the presence of Salmonella spp. and other enteric pathogens to monitor the prevalence of antimicrobial resistance among these pathogens. The NYSDOH developed a rapid and cost‐effective real‐PCR assay to screen for Salmonella spp. in retail meat products.

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“…However, an enrichment step is still required to detect small numbers of Salmonella in food samples. This step may consist of non-selective enrichment with buffered peptone water (BPW) and selective enrichment with Rappaport-Vassiliadis (RV) or tetrathionate [ 6 ]. These enrichment broths have been directly utilized for Salmonella DNA template preparation.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of PCR inhibitory effect of enrichment broths and comparison of DNA extraction methods for detection of Salmonella Enteritidis using real-time PCR assay

et al. 2010

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The best enrichment broth and DNA extraction scheme was determined for rapid and sensitive detection of Salmonella Enteritidis in steamed pork using real-time PCR. The inhibitory effect of commonly used Salmonella enrichment broths, Rappaport-Vassiliadis (RV) and Muller-Kauffmann tetrathionate with novobiocin (MKTTn), on real-time PCR was confirmed. The inhibition of PCR was statistically significant (p < 0.05) in RV and MKTTn, as compared with buffered peptone water (BPW) or phosphate-buffered saline. The inhibitory effect of the selective enrichment media was successfully removed by using a modified DNA extraction, PrepMan Ultra Reagent with an additional washing step or the DNeasy Tissue Kit. In three experiments, when applied to detection of Salmonella Enteritidis in steamed pork, the real-time PCR coupled with single 24 h enrichment with BPW performed better than double 48 h enrichment with BPW plus RV or MKTTn. The simple real-time PCR assay using BPW proved to be a rapid and sensitive test for detection of low concentrations of Salmonella Enteritidis in steamed pork samples as compared with the conventional culture method.

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Salmonella in Foods: New Enrichment Procedure for TECRA Salmonella Visual Immunoassay Using a Single RV(R10) Only, TT Only, or Dual RV(R10) and TT Selective Enrichment Broths (AOAC Official Method 998.09): Collaborative Study

Cited by 7 publications

References 0 publications

Toward the Adoption of Loop-Mediated Isothermal Amplification for Salmonella Screening at the National Antimicrobial Resistance Monitoring System's Retail Meat Sites

Toward the Adoption of Loop-Mediated Isothermal Amplification for Salmonella Screening at the National Antimicrobial Resistance Monitoring System's Retail Meat Sites

Detection ofSalmonellaspp. in retail meat products: a comparison between a discontinued commercial kit and a laboratory‐developed screening method

Evaluation of PCR inhibitory effect of enrichment broths and comparison of DNA extraction methods for detection of Salmonella Enteritidis using real-time PCR assay

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