中国人兽共患病学报2017，Vol.33Issue(1)：1-8,8.DOI:10.3969/j.issn.1002-2694.2017.01.001
医院临床标本中阴沟肠杆菌遗传多样性分析及应用TaqMan荧光定量PCR方法快速检测阴沟肠杆菌的研究
Genetic diversity of Enterobacter cloacae strains isolated from hospital in China and TaqMan probe real-time polymerase chain reaction for rapid detection of it
摘要
Abstract
Enterobacter cloacae are clinically important as nosocomial pathogens.In order to investigate the genetic diversity of the clinical E.cloacae,237 isolates obtained from routine diagnostic laboratory were examined with analysis of heat shock protein 60 gene (hsp60) sequence.Based on the neighbor-joining tree of the hsp60 gene sequence,ten genetic clusters of E.cloacae could be isolated from the clinical samples.Three genetic clusters (Ⅲ,Ⅵ and Ⅷ) represent almost 71％ of the isolates;cluster Ⅰ accounts for 11％;cluster Ⅵ,Ⅹ and Ⅻ were absent.The remaining six clusters are minority in our study,which totally accounted for 18％ of all strains.Based on out membrane protein Ⅹ (ompⅩ) gene sequence analysis of 237 strains,two sets of primers and probes were designed which were specific for ten clusters and cluster Ⅰ respectively.The limit of detections of the assay were 3.6 × 101 copies/μL for ten clusters and 2.1 × 101 copies/μL for cluster Ⅰ strains within 40 cycles.This method was also successfully applied to detect ten clusters and cluster Ⅰ strains from swab samples,the limit detection for swab samples with inoculated bacteria were 104 CFU/mL.In the study,we analyzed the genetic clusters of E.cloacae isolated from hospital setting,and developed a novel real-time polymerase chain reaction method for rapid detection of ten clusters and cluster Ⅰ.关键词
热休克蛋白60/TaqMan荧光定量PCR/外膜蛋白基因ⅩKey words
heat shock protein 60/TaqMan real-time polymerase chain reaction/out membrane protein Ⅹ分类
医药卫生引用本文复制引用
王怡倩,刘凯,李培京,熊衍文,叶长芸..医院临床标本中阴沟肠杆菌遗传多样性分析及应用TaqMan荧光定量PCR方法快速检测阴沟肠杆菌的研究[J].中国人兽共患病学报,2017,33(1):1-8,8.基金项目
We acknowledge the financial supports of the grants (Mega Project of Research on the Prevention and Control of HIV/AIDS,Viral Hepatitis Infectious Diseases 2011ZX10004-001,2013ZX10004-101 to Ye Chang-yun) from the Ministry of Science and Technology,China.国家重大传染病防治科技重大专项(No.2011ZX10004-001,No.2013ZX10004-101) （Mega Project of Research on the Prevention and Control of HIV/AIDS,Viral Hepatitis Infectious Diseases 2011ZX10004-001,2013ZX10004-101 to Ye Chang-yun）
