Establishment and preliminary application of a real-time fluorescent quantitative PCR assay for detection of feline herpesvirus 1
10.3969/j.issn.1671.7856.2014.012.010
- VernacularTitle:猫疱疹病毒Ⅰ型实时荧光定量 PCR方法的建立及初步应用
- Author:
Ji WANG
;
Rui FU
;
Wei LI
;
Xiaobo LI
;
Shujing WANG
;
Wei GONG
;
Bingfei YUE
;
Zhengming HE
- Publication Type:Journal Article
- Keywords:
Feline herpesvirus 1;
Real-time fluorescent quantitative PCR;
Cat
- From:
Chinese Journal of Comparative Medicine
2014;(12):47-54
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a real-time fluorescent quantitative PCR ( Q-PCR) method for detection of feline herpesvirus 1 ( FHV-1 ) in experiment cats and clinical sick cats.Methods Primers and TaqMan probes were designed and synthesized according to the published FHV-1 specific sequences of TK gene.FHV DNA standards were prepared using molecular biological techniques.The linearity, specificity, sensitivity, stability of the established Q-PCR method were tested.The method was used to detect 48 samples of cats.Results The linear range was 102 copies/μL to 109 copies/μL.The developed Q-PCR method showed no cross reaction with herpes virus type 1 ( HSV-1 ) , canine herpesvirus (CHV), pig pseudo rabies virus (PRV) and cat parvovirus (FPV).The sensitivity was 10 copies/μL.The coefficient of variation ( CV ) was less than 5%.There were 33 positive cases detected in the 48 samples of cats. Conclusions The developed Q-PCR method is good in linearity, specificity, sensitivity, stability, and may be used for rapid quantitative detection of FHV-1 in cats.
