Development of fluorescent quantitative one step RT-PCR method for the detection of GIV norovirus
10.3760/cma.j.issn.1003-9279.2016.02.024
- VernacularTitle:GIV诺如病毒荧光定量一步RT-PCR方法的建立
- Author:
Yuanyun AO
1
;
Jiemei YU
;
Dongmei ZHOU
;
Miao JIN
;
Lili LI
;
Zhaojun DUAN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所腹泻室
- Keywords:
norovirus;
GIV;
fluorescent quantity
- From:
Chinese Journal of Experimental and Clinical Virology
2016;30(2):208-211
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a fluorescent quantitative one step real time-PCR method for the detection of GIV NoVs.Methods Specific primers and TaqMan probe of GIV NoVs were analysed and synthesized.Then the reaction system and conditions were optimized and the RNA standard was conducted according to the sequence of human GIV NoVs of our nation.The sensitivity,specificity,and repeatability of this method were evaluated,and the traditional RT-PCR method was used to assess the method.Results The sensitivity of this method reached 1.0 × 102 copies/μl,and the detection range was 102 ～ 108copies/μl.No cross reaction with NoVs (GⅠ,GⅡ),enteric adenovirus,rotavirus,astrovirus and others was found,showing high specificity.By repeated experiments,the intra-assay and inter-assay coefficient of variation (CV) about Ct value was less than 1.31％ and 3.68％ respectively,which showed good repeatability.The method has high sensitivity and accurate quantity advantage by comparison with the traditional RT-PCR on detection of positive samples.Conslusions The developed fluorescent quantitative one step real time PCR method for identification of the GIV NoVs will be applied to detecting GIV NoVs in gastroenteritis patients.
