Diarrhoeagenic E. coli · Multiplex PCR
EPEC STEC EHEC Multiplex PCR Kit – identification of EPEC/STEC/EHEC
The EPEC STEC EHEC Multiplex PCR Kit identifies enteropathogenic, Shiga toxin-producing and enterohaemorrhagic E. coli by multiplex PCR. One reaction amplifies the virulence genes bfpB, escV, stx1, stx2 and uidA, and the band pattern on an agarose gel shows the pathotype.
- Multiplex end-point PCR
- Gel read-out on 1.5 % agarose
- Total assay within about 3 hours
How EPEC STEC EHEC Multiplex PCR Kit works
Dry-powder PCR tubes are reconstituted, lysed colony DNA is added and the reaction runs in a standard thermocycler. Band sizes on the gel are compared with the marker; the uidA band confirms E. coli, and the virulence gene bands give the pathotype.
Technical data
| Preparation | Dry-powder tubes + 18 µL PCR reconstitution solution; controls 80 µL ultrapure water; lysis 99 °C 10 min |
| Incubation | PCR 94 °C 5 min; 35 cycles (94 °C 30 s / 59 °C 30 s / 72 °C 90 s); 72 °C 5 min |
| Storage | 2-8 °C (reconstituted controls/DNA -20 °C) |
| Shelf life | 12 months |
Applications and practical notes
E. coli O157 and other Shiga toxin-producing E. coli (STEC) are tested in meat, raw milk products, sprouts and vegetables, for example according to ISO 16654 and FDA BAM Chapter 4. Typical applications include:
- Raw and minced meat, especially beef
- Raw milk and raw-milk cheese
- Sprouts, leafy greens and other fresh produce
- Outbreak investigations and environmental samples
Working with EPEC STEC EHEC Multiplex PCR Kit: Test pure colonies from MacConkey or EMB agar, and always run the negative and positive controls supplied.
Good practice with rapid tests: store the kit as stated on the label, let the components reach room temperature before use and use each item only once. Record the lot number and the result, and confirm positive results according to the reference method when this is required.
About coliform and E. coli methods: coliforms and E. coli are counted either by the multiple-tube (MPN) method, with a presumptive broth followed by confirmation broths, or by colony counts on selective agars. FDA BAM Chapter 4 describes both approaches for food and water, and ISO 16649-2 gives the colony-count method for β-glucuronidase-positive E. coli on TBX Agar. E. coli O157 and other STEC need dedicated media, serology and PCR. COLO.Science offers media, supplements, rapid tests, identification kits and QC strains for every step.
Complete sets from one supplier: COLO.Science supplies culture media, supplements, rapid tests, identification kits and QC strains as complete sets for the method you follow. Send us your sample types and standards, and we will prepare an offer with the matching products and documentation.
EPEC STEC EHEC Multiplex PCR Kit is supplied by COLO.Science in the pack sizes listed under Ordering information. For larger quantities, regular deliveries or method-specific sets that include EPEC STEC EHEC Multiplex PCR Kit, please send us a request. With every delivery of EPEC STEC EHEC Multiplex PCR Kit you receive the lot number and expiry date, and on request we send the available documentation for EPEC STEC EHEC Multiplex PCR Kit.
Where EPEC STEC EHEC Multiplex PCR Kit fits in E. coli testing
- Presumptive test – Lauryl Sulfate Tryptose Broth (MPN) or VRBL / chromogenic agar (colony count)
- Confirmation – BGLB Broth for coliforms; EC or EC-MUG Broth at 44.5 °C for E. coli
- Isolation – EMB, MacConkey, TBX or chromogenic E. coli agar
- Identification – IMViC kit; for O157 and STEC: SMAC or chromogenic O157 agar, latex and PCR
Ordering information
| Product | COLO code | Pack |
|---|---|---|
| EPEC STEC EHEC Multiplex PCR Kit | PRO7422 | 24 tests |
Product notice: preparation, storage conditions and shelf life are given on the label. Technical data on this page are for orientation; the label and the lot documentation apply. B2B prices are quoted net, excluding VAT.
Sales and technical support
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Quotations and technical support for culture media and microbiology products across our international sales territory. B2B prices are net, excluding VAT.
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Need the complete set for your method?
Send the sample type, target organism and the standard you follow. COLO.Science will put together the media, supplements, QC strains and confirmation tests for the whole procedure.
