Evaluation of the BAX® System for Detection of Salmonella in Selected Foods: Collaborative Study

Silbernagel, Karen M; Jechorek, Robert P.; Carver, Charles N; Barbour, W. Mark; Mrozinski, Peter

doi:10.1093/jaoac/86.6.1149

Cited by 27 publications

(6 citation statements)

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“…Furthermore, the quantification metrics were analyzed for main effect and interactions using a Mann-Whitney U test. Pairwise differences comparing significances between the time (16,18, and 20 h) of Campylobacter jejuni, coli, and lari and the species Campylobacter species (C. jejuni, C. coli, and C. lari) at 16, 18, and 20 h of enrichment were evaluated using an χ 2 Analysis. Significant differences were considered at p ≤ 0.05.…”

Section: Objective 2: Time Of Detection Of Campylobacter Spp In Enric...mentioning

confidence: 99%

“…To further delineate the effect of enrichment time on the PCR performance, the main effect and interaction of enrichment time (16,18, and 20 h) and Campylobacter species (C. jejuni, coli, and lari) was determined (Table 2). There was an interaction between the enrichment time and Campylobacter species on PCR performance metrics, sensitivity, NLR, NPV, and accuracy (p < 0.05; Table 2).…”

Section: Objective 2: Time Of Detection Of Campylobacter Spp In Enric...mentioning

confidence: 99%

“…Results of sensitivity, specificity positive likelihood ratio (PLR), negative likelihood ratio (NLR), prevalence, positive predictive value (PPV), negative predictive value (NPV), and accuracy of the BAX ® System Real-Time PCR assay for the detection of each Campylobacter species between media, enrichment times 16, 18, and 20 h in 2× blood-free Bolton broth (2× BFBB); Table S6. Statistical significance between the sensitivity, accuracy, prevalence, negative likelihood ratio (NLR), negative predictive value (NPV) between the time (16,18, and 20 h) of Campylobacter jejuni, coli, and lari; Table S7. Statistical significance between the sensitivity, accuracy, prevalence, negative likelihood ratio (NLR), negative predictive value (NPV) between Campylobacter spp., jejuni, coli, and lari, at 16, 18, and 20 h of enrichment; Table S8.…”

Section: Supplementary Materialsmentioning

confidence: 99%

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Adaptation of a Commercial Qualitative BAX® Real-Time PCR Assay to Quantify Campylobacter spp. in Whole Bird Carcass Rinses

Bodie,

Dittoe,

Applegate

et al. 2023

Foods

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Poultry is the primary reservoir of Campylobacter, a leading cause of gastroenteritis in the United States. Currently, the selective plating methodology using selective agars, Campy Cefex and Modified Charcoal Cefoperazone Deoxycholate agar, is preferentially used for the quantification of Campylobacter spp. among poultry products. Due to the specific nature of Campylobacter, this methodology is not sensitive, which can lead to skewed detection and quantification results. Therefore, Campylobacter detection and quantification methods are urgently needed. The objective was to develop a shortened enrichment-based quantification method for Campylobacter (CampyQuant™) in post-chill poultry rinsates using the BAX® System Real-Time PCR assay for Campylobacter. The specificity and sensitivity for the detection of C. jejuni, C. coli, and C. lari in pure culture were determined. The BAX® System Real-Time PCR assay consistently detected and identified each species 100% of the time with an enumeration range of 4.00 to 9.00 Log10 CFU/mL. Enrichment time parameters for low-level concentrations (0.00, 1.00, and 2.00 Log10 CFU/mL) of Campylobacter using the BAX® System Real-Time PCR assay were elucidated. It was determined that an enrichment time of 20 h was needed to detect at least 1.00 Log10 CFU/mL of Campylobacter spp. Using the BAX® System Real-Time PCR assay for Campylobacter. As a result, time of detection, detection limits, and enrichment parameters were used to develop the CampyQuant™ linear standard curve using the detected samples from the BAX® System Real-Time PCR assay to quantify the levels in post-chill poultry rinsates. A linear fit equation was generated for each Campylobacter species using the cycle threshold from the BAX® System Real-Time PCR assay to estimate a pre-enrichment of 1.00 to 4.00 Log10 CFU/mL of rinsates detected. The statistical analyses of each equation yielded an R2 of 0.93, 0.76, and 0.94 with a Log10 RMSE of 0.64, 1.09, and 0.81 from C. jejuni, C. coli, and C. lari, respectively. The study suggests that the BAX® System Real-Time PCR assay for Campylobacter is a more rapid, accurate, and efficient alternative method for Campylobacter enumeration.

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Section: Objective 2: Time Of Detection Of Campylobacter Spp In Enric...mentioning

confidence: 99%

Section: Objective 2: Time Of Detection Of Campylobacter Spp In Enric...mentioning

confidence: 99%

Section: Supplementary Materialsmentioning

confidence: 99%

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Adaptation of a Commercial Qualitative BAX® Real-Time PCR Assay to Quantify Campylobacter spp. in Whole Bird Carcass Rinses

Bodie,

Dittoe,

Applegate

et al. 2023

Foods

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“…A lysate sample was prepared from each regrowth sample according to the BAX ® Salmonella assay. The lysates were analyzed on the BAX ® Q7 instrument with the BAX ® Salmonella PCR kit (Cheung et al, 2007;D'Aoust et al, 2007;Koyuncu et al, 2010;Silbernagel et al, 2003;Tice et al, 2009). All regrowth samples that had positive BAX ® results were used to inoculate in the selective enrichment broth and then were processed through the IMS for further confirmation.…”

Section: Salmonella Analysis-the Bax ® Pcrmentioning

confidence: 99%

Detection and Isolation of Salmonella spp. in Animal Feeds from 2007-2011

Hsieh¹,

Lee

Poole

et al. 2014

regsci

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Salmonella species (spp.) are zoonotic pathogens that contaminate animal ingredients and finished feed and represent a significant hazard as identified by the Codex Animal Feed Taskforce. The United States (US) Food and Drug Administration (FDA) promulgated regulations prohibiting Salmonella contamination in feed and has since published a draft guidance document expressing their current strategy involving regulatory oversight of Salmonella contamination in feed. The Office of the Texas State Chemist (OTSC) initiated the broad surveillance of Salmonella spp. in 2007 in response of a Salmonella enterica serotype Typhimurium outbreak in frozen rodents, which are subject to the agency’s regulatory oversight as defined by the Texas Commercial Feed Control Act. From calendar years of 2007-2011, 2622 total feed samples were collected and subsequently evaluated for Salmonella contamination using multiple first-line methods, the accuracy of which was assessed with DNA-based polymerase chain reaction (PCR) and automated immunoanalysis. Three hundred and five out of 2622 tested samples with 78 different serotypes were identified as Salmonella-contaminated products. Results also indicated a steady increase in Salmonella incidence, along with a corresponding increase in Salmonella isolate and serotype diversity since 2007. This prevalence of Salmonella-contaminated feed stocks presents a potential risk to public health.

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“…Rapid molecular-based methodologies, such as real-time polymerase chain reaction (qPCR), are alternatives that can improve the accuracy and efficiency of SE testing and confirmation [13][14][15][16][17]. Unlike the traditional Salmonella culture-based approaches, several investigations have shown that qPCR methods provide superior efficiency and specificity in detecting SE in a variety of food matrices, including shell eggs [16,18,19].…”

Section: Introductionmentioning

confidence: 99%

A Real-Time PCR Approach for Rapid Detection of Viable Salmonella Enteritidis in Shell Eggs

Chan¹,

Liau²,

Low³

et al. 2023

Microorganisms

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Rapid and robust detection assays for Salmonella Enteritidis (SE) in shell eggs are essential to enable a quick testing turnaround time (TAT) at the earliest checkpoint and to ensure effective food safety control. Real-time polymerase chain reaction (qPCR) assays provide a workaround for the protracted lead times associated with conventional Salmonella diagnostic testing. However, DNA-based analysis cannot reliably discriminate between signals from viable and dead bacteria. We developed a strategy based on an SE qPCR assay that can be integrated into system testing to accelerate the detection of viable SE in egg-enriched cultures and verify the yielded SE isolates. The specificity of the assay was evaluated against 89 Salmonella strains, and SE was accurately identified in every instance. To define the indicator for a viable bacteria readout, viable or heat-inactivated SE were spiked into shell egg contents to generate post-enriched, artificially contaminated cultures to establish the quantification cycle (Cq) for viable SE. Our study has demonstrated that this technique could potentially be applied to accurately identify viable SE during the screening stage of naturally contaminated shell eggs following enrichment to provide an early alert, and that it consistently identified the serotypes of SE isolates in a shorter time than conventional testing.

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Evaluation of the BAX® System for Detection of Salmonella in Selected Foods: Collaborative Study

Cited by 27 publications

References 1 publication

Adaptation of a Commercial Qualitative BAX® Real-Time PCR Assay to Quantify Campylobacter spp. in Whole Bird Carcass Rinses

Adaptation of a Commercial Qualitative BAX® Real-Time PCR Assay to Quantify Campylobacter spp. in Whole Bird Carcass Rinses

Detection and Isolation of Salmonella spp. in Animal Feeds from 2007-2011

A Real-Time PCR Approach for Rapid Detection of Viable Salmonella Enteritidis in Shell Eggs

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