VanB表型-vanA基因型VRE分子特征及遗传背景研究

目的 探讨VanB表型-vanA基因型VRE耐药转座子结构、分子特征及遗传背景,并与VanA表型-vanA基因型VRE进行比较分析,以确定基因型与表型不一致的形成机制.方法 收集2008年3月至2009年1月卫生部北京医院临床标本中21株VRE菌株,用Etest法对10种抗生紊进行MIC测定,并通过PCR、序列测定、接合试验、耐药转座子结构、PFGE及MLST进行分子特征和遗传背景研究.结果 21株VRE均为vanA基因型,其中3株菌呈现VanB表型(万古霉素耐药,替考拉宁敏感);21株菌属于9个不同PFGE型,6个不同MIST型;多对引物对转座子的不同区域PCR扩增并进行序列拼接、比对,发现Tn1546结构中vanX、vanY的缺失及ISEfa4的插入与VanB表型-vanA基因型VRE菌株形成相关.结论 VanB表型-vanA基因型VRE菌株在国内较为罕见,Tn1546结构的改变与菌株的基因型与表型不一致相关.

The molecular characteristics and genetic background of VanB phenotype-van A genotype vancomycin resistant Enterococcus

Objective To investigate the structure of Tn1546 like elements,the molecular features and genetic background of VanB phenotype-vanA genotype VREs and to explain the difference between phenotype and genotype. Methods Twenty-one VREs were collected in Beijing Hospital from March 2008 to January 2009. Etest was used to determin MICs of ten antibiotics. PCR product sequencing, PFGE and MLST were performed to study the molecular features and genetic background of the 21 VREs. Results All VREs were vanA genotype, but 3 of them( 14. 3% ) exhibited the VanB phenotype. Based on PFGE analysis, 21 VREs belonged to 9 different patterns. Six STs were identified by MLST analysis. The analysis of the structure of Tn1546 like elements showed the deletion of vanY, vanZ and the insertion of ISEfa4 in orf2-vanR intergenic region may be related to the formation of VanB phenotype-wanA genotype. Conclutions VanB phenotype-vanA genotype VREs were rarely found in China. The results of vanA cluster rearrangements could partly explain the causes of difference between phenotype and genotype.