Establishment and application of Real-time quantitative PCR for human Metapneumovirus detection
10.3760/cma.j.issn.1003-9279.2016.02.026
- VernacularTitle:偏肺病毒逆转录实时荧光定量PCR的建立及应用
- Author:
Jieying ZHOU
1
;
Yingbiao LIN
;
Youde CAO
;
Zhaojun DUAN
Author Information
1. 湖南省郴州市第一人民医院
- Keywords:
Real-time quantitative reverse transcription PCR;
Metapneumovirus
- From:
Chinese Journal of Experimental and Clinical Virology
2016;30(2):216-219
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish and evaluate a Metapneumovirus real-time quantitative reverse transcription polymerase chain reaction (RT-PCR),detect clinical specimens and explore the clinical prevalence characteristics of Metapneumovirus-infected children.Methods The primers and probes which targeted the conserved genes F were designed,RNA standards were prepared to establish a standard curve,the sensitivity,specificity and reproducibility had been tested.222 lower respiratory clinical specimens were collected from children in Lanzhou area with ARI.Metapneumovirus and co-infection viruses were detected simultaneously;further Metapneumovirus related epidemiology was studied.Results Metapneumovirus linear detection range was 10-10s copies/μl,the lowest detection limit was 10 copies/μl,the correlation coefficient was 1,the amplification efficiency was 91.62％,the CV of Ct value was less than 2％.Take conventional PCR product sequence results as reference,real-time quantitative RT-PCR sensitivity and specificity were 100％ and 96.17％.Metapneumovirus detection rate were 9.46％,13 cases for boys (5.86％),8 case for girls (3.60％).The detection rate of spring,summer,autumn and winter were 15.71％,0％,5.08％,11.11％.There were no significant differences between the Metapneumovirus viral load and mixed infection or the types of disease,clinical symptoms.Conclusions Metapneumovirus realtime quantitative RT-PCR has been confirmed as a sensitive and specific method.Metapneumovirus was an important agent of children respiratory tract infection in Lanzhou region.We should take the long-term systematic surveillance seriously.
