USE OF PRIMERS SELECTIVE FOR VANCOMYCIN RESISTANCE GENES TO DETERMINE VAN GENOTYPE IN ENTEROCOCCI AND TO STUDY GENE ORGANIZATION IN VANA ISOLATES
USE OF PRIMERS SELECTIVE FOR VANCOMYCIN RESISTANCE GENES TO DETERMINE VAN GENOTYPE IN ENTEROCOCCI AND TO STUDY GENE ORGANIZATION IN VANA ISOLATES
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DOI：
10.1128/aac.39.8.1772
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发表时间：
1995-08-01
影响因子：
4.9
通讯作者：
GOLDSTEIN, BP
中科院分区：
文献类型：
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作者：
MIELE, A;BANDERA, M;GOLDSTEIN, BP
肠球菌对万古霉素的耐药性是一个新兴的治疗问题。标准微生物学方法并不总能检测出耐药性。设计用于PCR的寡核苷酸引物，用于扩增vanA集群基因的特定区域，以及vanB和vanCl。这些引物正确地鉴定了30株万古霉素耐药菌株（17株VanB、7株VanB和6株鸡肠球菌）。casseliflavus肠球菌和万古霉素敏感菌株未见扩增。通过PCR和Southern blotting分析发现，17株VanA分离株均具有相同序列的orf- 1、orf-2、vanR、vanS、vanH、VanA和vanY基因，且vanR- VanA片段的基因间距离相同。该方法适用于肠球菌中不同万古霉素耐药基因型的快速检测。
Vancomycin resistance in enterococci is an emerging therapeutic problem. Resistance is not always detected by standard microbiological methods. Oligonucleotide primers for PCR were designed to target amplification of defined regions of genes of the vanA cluster, as well as vanB and vanCl. These primers correctly identified 30 vancomycin-resistant isolates tested (17 VanA, 7 VanB, and 6 Enterococcus gallinarum). No amplification was observed with Enterococcus casseliflavus or vancomycin-susceptible strains. Using PCR and Southern blotting, we found that all 17 VanA isolates had orf-l, orf-2, vanR, vanS, vanH, vanA, and vanY genes in the same sequence and that the intergenic distances in the vanR-vanA segments were the same. The described methods should be applicable to the rapid detection of the different vancomycin resistance genotypes in enterococci.