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| Product Manual BactoReal® E. coli Typing (STEC) stx1 & stx2 |
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| Material Safety Data Sheet BactoReal® E. coli Typing (STEC) stx1 & stx2 |
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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).
Description: BactoReal® Typing Kit E. coli is a series of different real-time PCR kits for detection of virulence-associated genes of E. coli. Depending on which kit and which combinations of kits are used, they allow the typing of enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC) and of Shiga toxin (verotoxin) producing E. coli (STEC). Assays for stx1 and stx2, as well as assays for eae and ehxA can be combined in a multiplex PCR. All kits consists of an assay mix for the detection of the pathogen as well as a positive control and an internal positive control (IPC). To minimize PCR cross-contamination,the reaction mix included contains dUTP and uracil-N glycosylase (UNG).
- Amplification and detection: virulence-associated genes stx1 & stx2 of E. coli
- Real-time PCRwith primern, probes and positive control
- PCR- platforms: runs on all established standard real-time PCR- platforms
- Harmonized thermal profiles to run RNA and DNA samples simultaneously
PCR platforms: BactoReal® Typing Kit E. coli is developed and validated for the Applied Biosystems® 7500 System (Thermo Fisher Scientific), LightCycler® 480 (Roche) and Mx3005P® QPCR System (Agilent), but is also suitable for other real-time PCR instruments.