Detection of porcine E. coli virulence-associated genes (F4, F5, F6, F18, F41, STa1, STb2, LT, Stx2e, EAST1, fimA, AIDA-I, pAA, escV, cbf1, iucD, papC, pic). Typing of E. coli via melting curves
Background: Most types of diarrhea are caused by strains of enterotoxigenic E. coli (ETEC). The main virulence factors associated with ETEC in diarrhoea are enterotoxins and fimbrial adhesins. The fimbrial types F4 (K88) and F18 are commonly found in pathogenic E. coli isolated from weaned pigs. F18 is most commonly found in Shiga toxin-producing E. coli (STEC) and the Stx2e variant has been associated with oedema disease in pigs. The fimbrial types F5 (K99), F6 (987P) and F41 are more frequently associated with E. coli causing neonatal diarrhea. Another virulence factor of ETEC is EAST1, which has also been detected in E. coli of different pathogenic strains, such as ETEC, enteropathogenic E. coli (EPEC) and STEC from humans and animals.
Description: BactoTyping E. coli contains PCR primer sets for the amplification and detection of porcine E. coli virulence-associated genes. The method is based on an intercalating dye using real-time PCR. Amplification curve and gene specific melt curve at 515 nm indicate the amplification of E. coli specific genes. Screening for porcine E. coli adhesion and toxin genes requires picking of several bacterial colonies per plate. Master mix and E. coli positive control are included.
Genotyping of porcine E. coli virulence-associated genes:
- F4, F5, F6, F18, F41
- enterotoxins (STa1, STb2, LT, Stx2e, EAST1),
- other adhesins (fimA, AIDA-I, pAA, escV, cnf1, iucD, papC, pic)
- Real-time PCR and melt curve analysis with supplied master mix
- ROX™ dye as passive reference