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Detection of vancomycin resistance genes combined with typing of Enterococci by means of multiplex PCR amplification and multiple primer DNA sequencing
Author(s) -
Monstein HansJürg,
Johansson Yvonne,
Jonasson Jon
Publication year - 2000
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1034/j.1600-0463.2000.d01-7.x
Subject(s) - biology , multiplex polymerase chain reaction , genotype , primer (cosmetics) , polymerase chain reaction , multiplex , typing , microbiology and biotechnology , multilocus sequence typing , gene , genetics , chemistry , organic chemistry
A multiplex PCR assay for the detection of vancomycin resistance ( van ) genes in enterococci was established. Primers targeting the 16S rRNA gene were included in the reaction mixture. Multipleprimer DNA sequencing of the PCR products provided species identification through partial nucleotide sequences of 16S rRNA genes, as well as confirmation of the correct identification of vanA, vanB, vanC‐1 , and vanC‐2/3 genotypes. Thirty‐nine enterococcal clinical isolates and type strains were examined for the presence of vancomycin resistance determinants. Twelve other isolates from a clinical reference collection (some of them having vanA, vanB, vanC‐1 , or vanC‐2/3 genotypes) were used as controls. Hybridization and partial DNA sequence analysis of multiplex PCR products revealed that none of the clinical isolates had a vanA genotype and only one had a vanB genotype. vanC‐1 was found in three clinical isolates, and vanC‐2/3 in one. Results obtained with the reference and type strains were in agreement with earlier results.