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. Key words: Polyomavirus; Revers trans criptase polymerase chain reaction; quantitative PCR

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