Real-time fluorescent quantitative PCR assay for detection of human rhinovirus

VernacularTitle:鼻病毒逆转录实时荧光定量PCR方法的建立及应用
Author: Jieying ZHOU ¹ ; Yaping SUN ; Haiyan CAO ; Zhiping XIE ; Zhaojun DUAN ; Youde CAO
Author Information

1. 湖南师范大学第一附属医院
Keywords: Rhinovirus; Polymerase chain reaction
From: Chinese Journal of Experimental and Clinical Virology 2014;28(2):129-131
CountryChina
Language:Chinese
Abstract: Objective To establish the method for detecting human Rhinovirus (HRV) subtypes A,B,and C by reverse real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and to assay HRV nasopharyngeal samples of children with acute respiratory tract infections.Methods The specific primers and Taqman probes of targeted HRV gene were designed,RNA standards were prepared to establish a standard curve,the sensitivity,specificity and reproducibility has been tested,222 clinical respiratory specimens were retrospectively detected.Results The linear ranges of human Rhinovirus A,B,and C subtypes were 109-10copies/μl,109-10 copies/μl,109-103copies/μl respectively,correlation coefficients were 0.999,0.997,0.999.Variation within the group was less than 3％.The detection rate was 27.02％ by RT-PCR,the sensitivity and specificity were 100％ and 98.18％.Conclusion The RT-PCR method can simultaneously detect rhinovirus subtypes A,B,and C with good sensitivity,specificity and reproducibility.