mericon VTEC stx1/2 Kit
For real-time PCR detection of shigatoxin-producing Escherichia coli (VTEC stx1 and VTEC stx2) in food or animal feed
The mericon VTEC stx1/2 Kit is designed for the target-specific detection of detection of shigatoxin-producing Escherichia coli (VTEC stx1 and VTEC stx2) DNA in food, animal feed, and pharmaceutical products. The assay uses a real-time PCR-based protocol and forms part of the comprehensive QIAGEN food testing portfolio, which also features dedicated sample preparation kits and assays for ingredient authentication and GMO DNA detection. The mericon VTEC stx1/2 assay performs optimally on the Rotor-Gene Q, but has also been validated for block thermal cyclers.
The mericon VTEC stx1/2 Kit is intended for molecular biology applications in food, animal feed, and pharmaceutical product testing. This product is not intended for the diagnosis, prevention, or treatment of a disease.
The mericon VTEC stx1/2 Kit is for the specific detection of shigatoxin-producing Escherichia coli in enrichment cultures of food and animal feed samples. The assay can detect as few as 10 copies of shigatoxin-producing E. coli DNA in a reaction and exhibits high specificity. In an experiment using 2500 copies of the tested DNA, no cross-reactivity was observed with other pathogens (see the table "Results of mericon VTEC stx1/2 Kit cross-reactivity study").
Results of mericon VTEC stx1/2 Kit cross-reactivity study
The mericon VTEC stx1/2 Kit is designed for the qualitative detection of shigatoxin-producing Escherichia coli (VTEC stx1 and VTEC stx2) in food and animal feed samples after enrichment. VTEC is an abbreviation for verotoxin-producing Escherichia coli and is synonymous with STEC, or shigatoxin-producing Escherichia coli. This nomenclature results from the interchangeable names for the two toxin groups: shigatoxins 1 and 2 (stx 1 and stx 2) and verotoxins 1 and 2. Some E. coli strains are human pathogens and lead to gastroenteritis and, less commonly, to hemolytic-uremic syndrome (HUS). Transmission of bacteria occurs mainly by fecal-oral contamination of food, but may also be transmitted by contact with diseased persons.
Detection of pathogens using real-time PCR is based on the amplification of a specific region of the relevant pathogen genome. The amplified product is detected using target-specific fluorescent probes that bind to it. As the PCR product accumulates, there is an increased fluorescent signal from the bound probes. Monitoring the fluorescence intensities during the PCR run (i.e., in real time) allows the detection of the accumulating PCR product without having to re-open the reaction tubes afterward.
mericon PCR Assays include an internal control to control potential inhibition. This aids in the straightforward interpretation of the results (see the table "Summary of possible PCR outcomes").
The PCR primer set for each assay is highly specific and targets a unique and conserved region of the tested pathogen genome that has been bioinformatically and experimentally verified.
DNA is extracted from an enrichment culture of a sample of food or animal feed using a dedicated sample preparation kit. The mericon Bacteria Kit is for the isolation of bacterial DNA from Gram-negative bacteria. The mericon Bacteria Plus Kit is for the isolation of bacterial DNA from Gram-positive bacteria.
The isolated, pure bacterial DNA is assayed using a streamlined real-time PCR protocol that returns results in 73 minutes on QIAGEN's Rotor-Gene Q. The protocol has also been validated for use on other thermal block cyclers.
The results are displayed as easy-to-interpret PCR curves.
The mericon VTEC stx1/2 Kit is for real-time PCR-based detection of shigatoxin-producing Escherichia coli (VTEC stx1 and VTEC stx2) in food, animal feed, and pharmaceutical products.
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