BactoReal® E. coli Typing eae & ehxA (EHEC, EPEC)

 

BactoReal E coli Typing eae_ehxABakterien Assay Vet FAM + VIC

 

Background: Escherichia coli is a gram negative bacterium that is a member of the normal gut flora of humans and animals. Most E. coli strains are harmless, but some serotypes carry virulence factors which enable them to be pathogenic in target hosts. These pathogenic E. coli are implicated in a number of diseases including diarrhoea, septicaemia and urinary tract infections in humans and animals.
Enteropathogenic Escherichia coli (EPEC) produces an outer membrane protein called intimin, encoded by eae and mediating intestinal cell attachment. Intimin is a virulence factor (adhesin) of EPEC and EHEC strains. Furthermore, EPEC are classified into typical and atypical strains based on the presence of the plasmid E. coli adherence factor (EAF). Typical EPEC strains are rarely isolated from animals, and humans are the major natural reservoir. The reservoir for atypical EPEC can be both animals and humans. By definition all EPEC lack genes to produce shiga toxin (stx).
Shiga toxin (verotoxin) producing Escherichia coli (STEC) is recognized as an important human diarrheal pathogen. Swine and cattle play an important role as a carrier of STEC. The ability of STEC to cause illness is due to the production of Shiga toxins Stx1 and/or Stx2 (encoded by stx1 and stx2 genes).
Enterohaemorrhagic Escherichia coli (EHEC) can cause severe foodborne disease. EHEC is also positive for stx1 and/or stx2 but also produces either intimin (eae) or hemolysin (ehxA).

Description: BactoReal® E. coli Typing eae & ehxA (EHEC, EPEC) is a real-time PCR test for detection of the virulence-associated genes eae and ehxA of EPEC (only positive for eae; negative for ehxA and stx1, stx2) and EHEC (positive for eae or ehxA; also positive for stx1 and/or stx2). Eae is detected in FAM channel. EhxA is detected in VIC/HEX channel. Both assays can be combined in a multiplex PCR. This test belongs to a series of different real-time PCR tests for detection of virulence-associated genes of E. coli (BactoReal® E. coli Typing).

PCR-platforms: BactoReal® E. coli Typing eae & ehxA (EHEC, EPEC) is developed and validated for the ABI PRISM® 7500 instrument (Life Technologies), LightCycler® 480 (Roche) and Mx3005P® QPCR System (Agilent), but is also suitable for other real-time PCR instruments.

Also available as kitversion: BactoReal® Kit E. coli Typing eae & ehxA (EHEC, EPEC)

Product BactoReal® E. coli Typing eae & ehxA (EHEC, EPEC)
Order no. RTGV936
Unit 100 reactions
Technology Real-time PCR
Target gene eae and ehxA gene, detection in FAM and VIC/HEX channel
Content Assay (primers + probe) for eae
Assay (primers + probe) for ehxA
Positive Control eae
Positive Control ehxA
Amplification mix & IPC Not included, see product description
PCR-Platforms Evaluated for ABI PRISM® 7500, LightCycler® 480 and MX3005P. Assay can also be used with other real-time PCR instruments.

 

Internal Positive Control (IPC) ÔÇô not included, optional: For exclusion of false-negative interpretation of results caused by PCR inhibition an ingenetix internal positive control assay (ControlReal 3, or Internal Positive Control Assay3) can be optionally performed in a multiplex PCR, depending on the PCR-platform. See product description.
– Control
Real 3 (order no. RTGMCR-3) contains primers, a probe and an internal control PCR target included in the assay mix.
Internal Positive Control Assay 3 (order no. RTGMIPC-3) contains primers, a probe and an internal control PCR target in an extra tube (for control of DNA extraction and PCR inhibition).

Series BactoReal® E. coli Typing
Product name Assay(s) for Detectionchannel Order no.
BactoReal® E. coli Typing stx1 & stx2 (STEC) stx1 & stx2 FAM + VIC/HEX RTGV935
BactoReal® E. coli Typing eae & ehxA (EHEC, EPEC) eae & ehxA FAM + VIC/HEX RTGV936
BactoReal® E. coli Typing eae (EHEC or EPEC) eae FAM RTGV937
BactoReal® E. coli Typing ehxA (EHEC) ehxA VIC/HEX RTGV938

 

BactoReal® E. coli Typing is a series of different real-time PCR tests for detection of virulence-associated genes of E. coli. Depending on which assay and which combinations of assays are used, they allow the typing of enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC) and of Shiga toxin (verotoxin) producing E. coli (STEC).
Stx1 is detected in FAM channel. Stx2 is detected in VIC/HEX channel. Eae is detected in FAM channel. EhxA is detected in VIC/HEX channel. Assays for stx1 and stx2, as well as assays for eae and ehxA can be combined in a multiplex PCR.


Ingenetix ViroReal®, BactoReal® and ParoReal Assays detecting viral, bacterial and parasitic DNA are optimized to run under the same thermal cycling conditions and with the same amplification mix.

 

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