军事医学Issue(7)：514-518,5.DOI:10.7644/j.issn.1674-9960.2015.07.006
耐万古霉素肠球菌检测基因芯片方法的建立和验证
Gene chip technique in detection of vancomycin-resistant Enterococcus gene
摘要
Abstract
Objective To develop a detection method based on the technology of gene chips which can quickly distinguish genes of Enterococcus faecalis, E.faecium and vancomycin resistance.Methods Based on the specific gene ( ddl) sequences of two types of Enterococcus from GenBank, oligonucleotide probes which could detect and distinguish special genes and drug resistance genes ( vanA,vanB) of Enterococcus were designed and compounded.Then,the probes were dotted to modified slide.The target DNA fragments of vancomycin-resistant Enterococcus ( VRE) were labeled with biotin by multiple PCR amplification, and then hybridized with oligonucleotide probes on slide.The results were analyzed by portable imager.The multiple PCR system, hybridization reaction and condition of the chemiluminescence method were optimized before the specificity, sensitivity and reproducibility of the chip were evaluated.Results One universal primer, four specific primers, one universal probe and four specific probes were selected.This gene chip was demonstrated of high specificity and repeatability.The detection sensitivity was 103 CFU/ml.The gene chip detection results of 10 clinical samples were basically consistent with the drug sensitivity test ( 8/10 ) .Conclusion A gene chip technique for the detection of VRE is established successfully.It is possible to distinguish the type of VRE and detect the genetic phenotypes of drug resistance by gene chip technique.关键词
万古霉素/肠球菌,粪/万古霉素抗药性/芯片分析技术/耐万古霉素肠球菌/多重PCR/化学发光法Key words
vancomycin/Enterococcus faecalis/vancomycin resistance/microchip analytical procedures/vancomycin-resistant Enterococcus/multiple PCR/chemiluminescence分类
医药卫生引用本文复制引用
何莎,宋燚,陈苏红,王升启,张五星,周伟,刘琪琦..耐万古霉素肠球菌检测基因芯片方法的建立和验证[J].军事医学,2015,(7):514-518,5.基金项目
“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项资助项目 （）
