湖南农业大学学报（自然科学版）2017，Vol.43Issue(3)：310-314,5.DOI:10.13331/j.cnki.jhau.2017.03.016
锦鲤疱疹病毒潜伏感染实时荧光定量PCR检测方法的建立
Method of Real-Time PCR for the detecting latent infection of koi herpesvirus
摘要
Abstract
To establish a rapid, sensitive and specific approach for detecting the latent infection of koi Herpesvirus (KHV), a Real-Time PCR test was designed through specific primers designed to target KHV Sph gene. Under the optimized reaction conditions, the results showed that this method was specific for latent detection of KHV without cross reaction to other fish pathogenic viruses, including carp pox herpes virus (CyHV-1), goldfish herpes virus (CyHV-2) and eel herpes virus (AngHV-1). The lowest limitation of detection concentration by Real-Time PCR was 42 copies per 1μL. A total of 63 clinical samples were detected by Real-Time PCR. The result showed that 18 samples out of which had koi herpesvirus disease (KHVD) history, all samples were positive; 15 samples out of which had in clinical signs and the positive rate was 100%; 30 samples out of which had no any clinical symptoms, and 6 out of the 30 samples were positive, the positive rate was 20%. In conclusion, the approach provides a rapid, sensitive and reliable molecular tool for detecting latent infection of koi Herpesvirus.关键词
锦鲤疱疹病毒(KHV)/潜伏感染/实时荧光定量PCR(Real-TimePCR)Key words
koi herpesvirus (KHV)/latent infection/Real-Time PCR (RT-PCR)分类
农业科技引用本文复制引用
陈俊杰,李媛媛,阳瑞雪,古晶晶,汪开毓,耿毅,欧阳萍..锦鲤疱疹病毒潜伏感染实时荧光定量PCR检测方法的建立[J].湖南农业大学学报（自然科学版）,2017,43(3):310-314,5.基金项目
中国博士后科学基金(2015M582563) （2015M582563）
教育部留学回国人员科研启动基金(教外司留[2015]311号) （教外司留[2015]311号）
