人多瘤病毒7 TaqMan探针实时定量PCR检测方法的建立  

Development of TaqMan Probe Real-Time Quantitation PCR Assay for Detection of Huamn Polyomavirus 7

作　　者：麻粉莲[1] 郑文芝[1] 魏田力[2] 张骞[1] 崔红[2] 郑丽舒[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052 [2]首都医科大学附属北京友谊医院儿科,北京100052

出　　处：《生物技术通讯》2015年第2期245-248,共4页Letters in Biotechnology

基　　金：国家传染病防治科技重大专项(2013ZX10004-101)

摘　　要：目的：建立人多瘤病毒7（HPyV7）核酸快速、特异的TaqMan探针实时定量PCR检测方法。方法：分别设计HPyV7特异性的引物与TaqMan探针，建立实时荧光定量PCR方法，并对其特异性、灵敏性和重复性进行评价；用建立的方法检测200份健康成人志愿者的血清和300份急性呼吸道感染住院儿童的鼻咽抽吸物样本。结果：所建立的实时定量PCR方法对HPyV7的检测灵敏度可达10拷贝/μL，检测线性范围为101-1010拷贝/μL，且实验特异性和重复性好（CV〈1.0%）；采用该方法，从200份血清样本中检出9份阳性标本。结论：建立了HPyV7 TaqMan探针实时定量PCR检测方法，为HPyV7的流行病学调查及其初步研究提供了技术手段。Objective： To develop a rapid and sensitive TaqMan based, real-time quantitation PCR（qPCR） assay for detection and quantitation of huamn polyomavirus 7（HPyV7）. Methods： The specific primers and TaqMan probe to develop a real-time qPCR assay for detection of HPyV7 were designed. The specificity, sensitivity and reproducibility of real-time qPCR were evaluated. Subsequently, real-time qPCR was used to identify HPyV7 in sera samples of 200 healthy adult volunteers and 300 nasopharyngeal aspirates of children with related pathogens of acute respiratory tract infections. Results： The analytical detection limit of this real-time qPCR assay was 10 copies/μL. This assay allowed quantitation of HPyV7 over span ranging from 101-1010 copies/μL, and showed good specificity and repeatability（CV〈1.0%）. A total of 200 sera specimens were screened for the presence of HPyV7 by using real-time qPCR and identified 9 specimens positive for HPyV7. Conclusion： A real-time qPCR assay for detection and quantitation of HPyV7 has been developed. This assay provides technical approach for an epidemiological investigation and preliminary study of HPyV7.

关 键 词：人多瘤病毒7 TAQMAN探针 实时定量PCR 

分 类 号：R373[医药卫生—病原生物学]