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Identification of Enterococcus spp. of the human normal microbiota by pyrosequencing

Abstract number: 902_p946

Lindgren M.

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Objectives:

The main objective was to develop a pyrosequencing method for identification of Enterococcus spp. species with Pyrosequencing method. Also, development of antibiotic resistance with special reference to macrolide resistance will be studied by susceptibility testing in samples isolated serially from subject exposed to clindamycin.

Methods:

Biochemical identification of the enterococcal strains from faecal samples was done by growth at 45°C, catalase and hydrolyse of 1-pyrridonyl-beta-naphtylamide (PYR). Species identification was done with Pyrosequencing method. PSQ 96MA pyrosequencing technique enabled identification of different Enterococcus species based on their 16S rRNA V2-regions signature-sequences. Antibiotic susceptibility testing was done by agar dilution method on Müller–Hinton II medium, according to NCCLS. MIC values were tested against erythromycin, clindamycin, ciprofloxacin, ampicillin, gentamicin, vancomycin and tetracycline. Macrolide resistance genes; erm(B), erm(TR) and mef(A) was studied by Multiplex-PCR.

Results:

With Pyrosequencing method, we identified 46 Enterococcus faecium, 22 E. faecalis, 11 E. avium and 33 E. casseliflavus species, and 54 non-enterococci species. The antibiotic susceptibility testing showed that 26.5% of the Enterococcus strains were resistant to erythromycin, 14.8% to ciprofloxacin and 17.4% to tetracycline. About 31.6% of the Enterococcae had erm(B)-gene.

Conclusion:

Pyrosequencing was rapid and easy method for identification of bacterial strains even to the species level. Antibiotic resistance varied a lot between different bacterial strains, as E. faecium and E. casseliflavus species being the most resistant ones. Pyrosequencing results correlated well with species phenotype and antibiotic resistance.

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Session Details

Date: 01/08/2007
Time: 00:00-00:00
Session name: XXIst ISTH Congress
Subject:
Location: Oxford, UK
Presentation type:
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