Detection and identification of human metapneumovirus by real time reverse transcription PCR
10.3760/cma.j.issn.1003-9279.2008.06.037
- VernacularTitle:实时荧光逆转录聚合酶链反应检测人类偏肺病毒方法的建立与应用
- Author:
Jian-Ming XING
1
;
Xue-Jun WENG
;
Su ZHANG
;
Xin-Hua YUAN
;
Cui-Fen SHEN
;
Ya-Qin ZHANG
;
Hong-Ling CHENG
;
Gang LI
;
Li-Hui YAO
Author Information
1. 湖州市妇幼保健院
- Keywords:
Real-time reverse transcriptase polymerase chain reactyion;
Metapneumovirus
- From:
Chinese Journal of Experimental and Clinical Virology
2008;22(6):510-512
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a rapid,sensitive and specific real time reverse transcription PCR for detecting and identifying human metapneumovirus. Methods The Hmpv-L gene of human metapneumovirus was chosen as target gene,the primers and TaqMan probe were designed,and the PCR reaction was optimized systematically. The total RNA was extracted from respiratory specimens,and reverse transcription was performed through random primer. The cDNA was detected by using real time PCR. The specificity,sensitivity and reproducibility of real time PCR were estimated. The real time PCR was applied to detect 180 clinical respiratory specimens. Results The human metapneumovirus can be detected using real time reverse transcription PCR accurately and quickly,and the sensitivity was 1 copy/μl.The coefficient of variation of intra-assay and inter-assay wasless than 5%. Among those 180 specimens,28(15.56%) were positive for human metapneumovirus,the clinical diagnoses for these 28 patients were pneumonia ( 15.60%,17/109) and bronehiolitis ( 15.49%,11/71 ) .21 positive specimens were from patients under 2 years of age,and 6 positive specimens were from patients between 2 and 5 years of age,only 1 positive specimens was from patients over 5 years. Conclusion It is demonstrated that real time reverse transcription PCR is a reliable,accurate and feasible assay for human metapneumovirus,whieh has become one of the most important pathogens induced acute respiratory infections in pediatric patients.
