Species identification and detection of vancomycin resistance genes in enterococci of animal origin by multiplex PCR

• Published in: International journal of food microbiology 2003-12, Vol.88 (2), p.305-309
• Main Authors: Mac, Kiem, Wichmann-Schauer, Heidi, Peters, Jan, Ellerbroek, Lüppo
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Cattle; Chickens; DNA, Bacterial - analysis; Enterococcus - classification; Enterococcus - drug effects; Enterococcus - genetics; Enterococcus faecalis; Enterococcus faecalis - classification; Enterococcus faecalis - drug effects; Enterococcus faecalis - genetics; Enterococcus faecium; Enterococcus faecium - classification; Enterococcus faecium - drug effects; Enterococcus faecium - genetics; Food industries; Food Microbiology; Fundamental and applied biological sciences. Psychology; General aspects; Methods of analysis, processing and quality control, regulation, standards; Multiplex PCR assay; PCR parameter; Polymerase Chain Reaction - methods; Sensitivity and Specificity; Species Specificity; Swine; vanA gene; vanB gene; vanC1 gene; vanC2 gene; Vancomycin Resistance - genetics; Vancomycin resistant enterococci
• ISSN: 0168-1605; 1879-3460
• DOI: 10.1016/S0168-1605(03)00192-2

A multiplex PCR assay for the differentiation of species and detection of vancomycin ( van) resistance genes in enterococci was established. Three hundred sixty-seven enterococcal strains isolated from food were selected from a sample collection and were examined with regards to the existence of four vancomycin resistance genes ( vanA, vanB, vanC1 and vanC2) and to species differentiation ( E. faecalis, E. faecium, E. casseliflavus and E. gallinarum) by multiplex PCR. Apart from unambiguous results for the majority of strains, the E. faecium specific primers used here, showed weak points in the specificity and sensitivity, respectively. Despite these rare instances, we succeeded in optimising the multiplex PCR assay by variation of annealing temperature and primer combination. The assay presented here, with its optimised parameters, is therefore suitable for routine use and because of time and material saving, it is an alternative and rapid method in comparison to conventional tests.