应用PCR-荧光探针法快速检测耐多药结核分枝杆菌基因型  

PCR-fluorescent probe in rapid detection of genotypes of multidrug-resistant Mycobacterium tuberculosis

作　　者：梁建琴[1] 李洪敏[1] 高华方[2] 刘莹莹[2] 王金河[1] 武丽红[1] 冯士生[1] 刘景阳[1] 史晓朋[1] 

机构地区：[1]解放军第309医院,北京100091 [2]博奥生物有限公司暨生物芯片北京国家工程研究中心,北京100084

出　　处：《中华医院感染学杂志》2013年第21期5140-5142,共3页Chinese Journal of Nosocomiology

基　　金："十一五"国家科技重大专项基金资助项目(2008ZX10003-001)

摘　　要：目的评价PCR-荧光探针法快速检测耐多药结核分枝杆菌基因型的临床应用价值。方法应用PCR-荧光探针法对142份涂阳痰液样本进行分子菌种鉴定,对鉴定为结核分枝杆菌复合群的标本进行katG、inhA、rpoB耐药基因突变位点检测,并与传统药敏试验结果比较。结果 142份涂阳痰样本中3份鉴定为非结核分枝杆菌,139份鉴定为结核分枝杆菌复合群;139株结核分枝杆菌中92株未检测出katG、inhA、rpoB基因耐药检测位点突变,其中传统药敏结果72株为HSRS、6株为HSRR、14株为HRRR;33株检测出katG和rpoB或inhA和rpoB两个基因耐药位点突变,传统药敏结果均为HRRR;11株检测出katG或inhA耐药基因位点突变,传统药敏结果均为HRRS;1株检测出rpoB耐药基因位点突变,传统药敏结果为HSRR,药敏试验符合率为84.17%;对部分样本行PCR产物直接测序(PCR-DS),结果显示,PCR-荧光探针和PCR-DS检测耐多药结核分枝杆菌基因型结果符合率为100.00%。结论 PCR-荧光探针法快速检测耐多药结核分枝杆菌基因型简便、快速,具有临床推广应用价值。OBJECTIVE To evaluate the clinical value of PCR-fluorescent probe in rapid detection of genotypes of multidrug-resistant Mycobacteriurn tuberculosis. METHODS The molecular bacterial identification of 142 smearpositive sputum specimens was performed with the use of PCR-fluorescent probe method, then the mutations of the drug resistance genes including katG, inhA, and rpoB were detected in the isolates identified as the Mycobacterium tuberculosis, and the drug susceptibility testing was carried out. RESULTS Of 142 smear-positive sputum specimens, 3 were identified as the non-Mycobacterium tuberculosis, 139 as the Mycobacteriurn tuberculosis. The mutations of the katG, inhA, and rpoB genes have not been detected in 92 of 139 strains of Mycobacterium tuberculosis. The result of the drug susceptibility testing indicated that there were 72 strains of HSRS, 6 strains of HSRR, and 14 strains of HRRR; the mutations in katG and rpoB genes or inhA and rpoB genes have been detected in 33 strains, all of which were identified as HRRR; the mutation of katG or inhA has been detected in 11 strains, all of which were identified as the HRRS; the mutation of rpoB has been detected in 1 strain, which was identified as the HSRR; the accordance rate of the drug susceptibility testing was 84.17%. The result of PCR-direct sequencing（PCR-DS） for the products revealed that the accordance rate of the PCR-fluorescent probe and the PCR-DS in detection of genotypes of multidrug-resistant Mycobacteriurn tuberculosis was 100.00 %. CONCLUSION The PCR-fluorescent probe technique is simple and rapid in detection of genotypes of multidrug-resistant Mycobacterium tuberculosis and is worthy to be promoted in the hospital.

关 键 词：PCR-荧光探针法 耐多药结核分枝杆菌 KATG基因 inhA基因 RPOB基因 

分 类 号：R378.911[医药卫生—病原生物学]