Development and application of TaqMan probe real-time PCR assay for detection of KIPyV and WUPyV
10.3760/cma.j.issn.1003-9279.2015.03.024
- VernacularTitle:人多瘤病毒KI和WU PCR检测方法的建立及应用
- Author:
Qian ZHANG
1
;
Wenzhi ZHENG
;
Wumei YUAN
;
Fenlian MA
;
Lishu ZHENG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所卫生部医学病毒和病毒病重点实验室
- Keywords:
Polyomavirus;
Revers trans criptase polymerase chain reaction;
quantitative PCR
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(3):266-269
- CountryChina
- Language:Chinese
-
Abstract:
Objective We develop a rapid,specific,sensitive tandardized SOP.And initial application for 200 nasopharyngeal aspirates of children with related pathogens of acute respiratory tract infections in BeiJing area.Methods To developed nested PCR and TaqMan probe real-time fluorescence quantitative PCR method for detection of KIPyV and WUPyV'S gene,and then the sequences of gene fragments are analyzed.Evalution of two assays from 200 nasopharyngeal aspirates.Results In this study,sensitivity of TaqMan probe real time fluorescent quantitative PCR assay was higher than one of nested PCR (500 copies/μl),and both assays did not show any positive amplification in detetion of other respiratory virus.Coefficient of varience of KIPyV and WUPyV are less than 2.9％ and 1.95％ respectively in the repeatability detection.The detection rates of KIPyV,and WUPyV were 1.5％ and 8％ in nested PCR assay and 12％ and 14％ in real time Fluorescent quantitative PCR assay respectively.Conclusion This study established good sensitivity and reproducibility,high specificity and rapid method for detection nucleic acid of these polyomaviruses that have good prospects on the clinical application.
