Monstein H J, Johansson Y, Jonasson J
Division of Clinical Microbiology, Molecular Biology Laboratory-LMO, Faculty of Health Sciences, University Hospital, Linköping, Sweden.
APMIS. 2000 Jan;108(1):67-73. doi: 10.1034/j.1600-0463.2000.d01-7.x.
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. Multiple-primer 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.
建立了一种用于检测肠球菌中万古霉素耐药（van）基因的多重PCR检测方法。反应混合物中包含靶向16S rRNA基因的引物。对PCR产物进行多引物DNA测序，通过16S rRNA基因的部分核苷酸序列进行菌种鉴定，并确认vanA、vanB、vanC-1和vanC-2/3基因型的正确鉴定。检测了39株肠球菌临床分离株和标准菌株中万古霉素耐药决定簇的存在情况。另外从临床参考菌株库中选取12株分离株（其中一些具有vanA、vanB、vanC-1或vanC-2/3基因型）作为对照。多重PCR产物的杂交和部分DNA序列分析表明，临床分离株中没有一株具有vanA基因型，只有一株具有vanB基因型。在3株临床分离株中发现了vanC-1，1株中发现了vanC-2/3。参考菌株和标准菌株的检测结果与早期结果一致。
