Detection of Vancomycin-resistant Enterococci, an evaluation of direct analyzing from ESwab using real-time PCR detection kit and culture

Sammanfattning: Background: Vancomycin-resistant enterococci (VRE) is a common nosocomial infection. It classifies as Enterococcus faecalis or faecium carrying vanA or vanB gene that alters the bacterial cell wall hence lowering affinity for Vancomycin. Screening for VRE in Swedish hospitals are performed with stool sample pre-grown in selective broth followed by PCR and culture on selective media. Viasures Vancomycin resistance, Real Time PCR Detection Kit indicates that pre-growth in broth is not needed for the analyze. Aim: Comparison between the PCR kit and the subsequent culture on chromogenic agar with or without pre-growth in selective broth. Method: E. faecium with vanA gen (CCUG 36804) and E. faecalis with vanB gen (ATCC 51299) were suspended in different concentrations and added to ESwab transport medium. Thereafter small samples from the ESwab tube were enriched in selective broth. Samples from both selective broth and ESwab medium were analyzed with Viasures PCR kit on BD-MAX system and cultivated on chromogenic agar. Results: With pre-growth in selective broth the genes were found in every sample regardless of pre-concentration in the ESwab medium. Without enrichment the PCR kit always amplified the genes when the concentration was 40 000 cfu/ml for E. faecium (vanA) and ≥ 10 000 cfu/ml for E. faecalis (vanB). Colonies grew on chromogenic agar in every concentration from both ESwab and selective broth. Conclusion: Culture on chromogenic agar is comparable with or without pre-growth in selective broth but Viasure’s PCR kit is not equal for both methods in lower concentrations of the bacteria.