荧光定量PCR检测查菲埃立克体  

DEVELOPMENT OF A QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION ASSAY SPECIFIC FOR EHRLICHIAE CHAFFEENSIS

作　　者：张晶波[1] 温博海[1] 陈梅玲[1] 杨晓[1] 李丽莉[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室

出　　处：《寄生虫与医学昆虫学报》2006年第4期212-216,共5页Acta Parasitologica et Medica Entomologica Sinica

基　　金：国家科技攻关项目(2003BA712A04-07)

摘　　要：依据查菲埃立克体16S rRNA基因序列设计特异性引物和TaqMan-MGB探针，以克隆的查菲埃立克体16S rRNA基因片段作DNA模板，建立实时荧光定量PCR检测方法。与套式PCR相比较，荧光定量PCR检测的灵敏度是其30倍；用荧光定量PCR检测其他相关立克次体和细菌DNA样本，检出结果为0；对荧光定量PCR检测重复性进行分析，变异系数（CV）批内和批间误差在0．2％～2．0％之间。结果证明本研究建立的荧光定量PCR方法具有种特异性和良好的重复性，可用于检测感染样本中的微量查菲埃立克体DNA。According to the 165 rRNA gene sequences specific for Ehrlichiae chaffeensis, a pair of primers and one TaqMan-MGB probe were designed. The real-time quantitative polymerase chain reaction was developed with the primers, probe, and the template （16S rRNA gene DNA fragment of E. chaffeensis）. The sensitivity of this real-time quantitative PCR was about 30 times higher than that of the nested PCR used to detect homologous DNA. By this quantitative real-time PCR, the DNA sample of E. chaffeensis was positively detected but not other rickettsial or bacterial DNA samples and the variation coefficients of intra- and inter-assay reproducibility were 0.2%～2.0%. The results show that the method is useful for detection of tiny DNA of E. chaffeensis in various samples.

关 键 词：查菲埃立克体 实时荧光定量PCR 16S RRNA基因 埃立克体感染 

分 类 号：R450[医药卫生—治疗学]