Comparison of Assurance GDS® MPX ID for Top STEC with Reference Culture Methods for the Detection of E. coli Top 6 STEC; Direct Confirmation of Top 6 STEC from Isolation Plates and Determination of Equivalence of PickPen® and FSIS OctoMACS™ Concentration Protocols

Feldsine, Philip T; Lienau, Andrew H; Shah, Khyati; Immermann, Amy L; Soliven, Khanh; Kaur, Mandeep; Kerr, David E.; Jucker, Markus; Hammack, Tom; Brodsky, Michael; Agin, James

doi:10.5740/jaoacint.15-0261

Cited by 6 publications

(3 citation statements)

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“…However, our results showed that Assurance® GDS method (using m-Ehec) and ISO/TS13136 (using BPW) allowed su cient growth of STEC to allow detection and con rmation in both vegetable and meat, with respectively 375 g and 25 g sample size tested. The results obtained with Assurance® GDS and the ISO/TS13136 method are consistent with previously for the detection of STEC serogroups in vegetables (Costa et al 2019;Feldsine et al 2016).…”

Section: Discussionsupporting

confidence: 90%

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Validation of larger test portion sizes for Escherichia coli Shiga toxin producer (STEC) detection by Immunomagnetic separation and Real Time PCR in meat and vegetables

Bouvier

Canizares

Hamadou³

et al. 2023

Preprint

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The aims of this study were (i) to evaluate the performance of the Assurance® GDS method combining immunomagnetic separation and Real Time PCR for STEC detection in pooled samples (up to 375g) of vegetables and meat and (ii) to compare its performances to that of the reference method ISO/TS-13136:2012 (25 g sample size) in artificially contaminated samples. The alternative Assurance® GDS method includes enrichment in proprietary broth at 41.5ºC for 10 h followed by primary screening of TOP 7 STEC containing eae, stx genes and O157:H7 markers using MPX-Top 7 (IMS+PCR), secondary screening for serogroup identification using MPX-ID/EHEC-ID (IMS+PCR). And finally, cultural confirmation from same IMS beads on two selective agars is set up. For reference method, the enrichment was performed in BPW at 41.5ºC for 18 h. A total of 120 samples of meat and vegetables, including 20 uninoculated and 100 samples spiked with stressed cells (<12 CFU/25 or 375g) of STEC, were analyzed using both methods. Our results showed that the Assurance® GDS method LOD50 ranged from 0.12 CFU/375 g to 0.522 CFU/375 g and the LOD95 from 0.518 CFU/375 g to 2.257 CFU/375 g. These data are similar to the LOD50 data of the reference method.

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Section: Discussionsupporting

confidence: 90%

“…This may result in greater di culty in isolating target bacteria from others on the agar. (2) A few authors have also described differences between serogroups isolation (Feldsine et al 2016;Fratamico et al 2011;Hallewell et al 2017).…”

Section: Discussionmentioning

confidence: 99%

Validation of larger test portion sizes for Escherichia coli Shiga toxin producer (STEC) detection by Immunomagnetic separation and Real Time PCR in meat and vegetables

Bouvier

Canizares

Hamadou³

et al. 2023

Preprint

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“…Many EHEC isolates produce this protein, including some that carry plasmids only distantly related to pO157 (Beutin et al, 1989). Accordingly, the production of enterohaemolysin can be used as a diagnostic marker of EHEC (Feldsine et al, 2016). Interestingly, enterohaemolysin is also produced by some LEE-positive, Shiga toxin-negative strains of E. coli obtained from cattle and sheep (Cookson et al, 2007).…”

Section: Dec Pathotypesmentioning

confidence: 99%

Are Escherichia coli Pathotypes Still Relevant in the Era of Whole-Genome Sequencing?

Robins-Browne

Holt

Ingle³

et al. 2016

Front. Cell. Infect. Microbiol.

130

120

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The empirical and pragmatic nature of diagnostic microbiology has given rise to several different schemes to subtype E.coli, including biotyping, serotyping, and pathotyping. These schemes have proved invaluable in identifying and tracking outbreaks, and for prognostication in individual cases of infection, but they are imprecise and potentially misleading due to the malleability and continuous evolution of E. coli. Whole genome sequencing can be used to accurately determine E. coli subtypes that are based on allelic variation or differences in gene content, such as serotyping and pathotyping. Whole genome sequencing also provides information about single nucleotide polymorphisms in the core genome of E. coli, which form the basis of sequence typing, and is more reliable than other systems for tracking the evolution and spread of individual strains. A typing scheme for E. coli based on genome sequences that includes elements of both the core and accessory genomes, should reduce typing anomalies and promote understanding of how different varieties of E. coli spread and cause disease. Such a scheme could also define pathotypes more precisely than current methods.

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Detection Methods for Shiga Toxins and Shiga Toxin-Producing E. coli

Silva¹,

Brandon²,

Skinner³

et al. 2017

Shiga Toxins

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Comparison of Assurance GDS® MPX ID for Top STEC with Reference Culture Methods for the Detection of E. coli Top 6 STEC; Direct Confirmation of Top 6 STEC from Isolation Plates and Determination of Equivalence of PickPen® and FSIS OctoMACS™ Concentration Protocols

Cited by 6 publications

References 0 publications

Validation of larger test portion sizes for Escherichia coli Shiga toxin producer (STEC) detection by Immunomagnetic separation and Real Time PCR in meat and vegetables

Validation of larger test portion sizes for Escherichia coli Shiga toxin producer (STEC) detection by Immunomagnetic separation and Real Time PCR in meat and vegetables

Are Escherichia coli Pathotypes Still Relevant in the Era of Whole-Genome Sequencing?

Detection Methods for Shiga Toxins and Shiga Toxin-Producing E. coli

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