鸭乙型肝炎病毒荧光定量PCR检测方法的建立及应用  

Development and Application of a Real-time PCR Assay for Detection of Duck Hepatitis B Virus

作　　者：盛媛 朱蕴暖 刘娜 刘存霞[1] 于可响[1] 胡峰[1] 郭效珍 李玉峰[1] 刘丽萍 马秀丽[1] 黄兵[1] SHENG Yuan;ZHU Yunnuan;LIU Na;Liu Cunxia;YU Kexiang;HU Feng;GUO Xiaozhen;LI Yufeng;LIU Liping;MA Xiuli;HUANG Bing(Shandong Institute of Poultry Science,Poultry Breeding Engineering Technology Center of Shandong Province,The Key lab of poultry disease diagnosis and immunology of Shandong Province,Shandong Academy of Agricultural Science,Jinan 250023,China)

机构地区：[1]山东省农业科学院家禽研究所,山东省禽病诊断与免疫重点实验室,山东省家禽育种工程技术研究中心,济南250023

出　　处：《中国动物传染病学报》2021年第5期31-35,共5页Chinese Journal of Animal Infectious Diseases

基　　金：山东省重点研发计划项目(2019GNC106044);山东省现代农业产业技术体系项目(SDAIT-11-01);山东省农科院农业科技创新工程(CXGX2021A39);山东省农业科学院农业科技创新工程学科团队专项经费(CXGX2018E11)。

摘　　要：本研究根据鸭乙型肝炎病毒(DHBV)基因组C基因序列设计一对特异性引物,建立一种快速检测DHBV的SYBR Green荧光定量PCR方法。该方法在7.8×10^(2)copies/μL~7.8×10^(8)copies/μL模板量时,呈现良好的线性关系,相关系数R 2为0.999。利用该方法对来自山东不同地区的95份临床样品进行检测。结果显示,该方法特异性好,临床样品检测阳性率与常规PCR方法的符合率为97.9%,且灵敏度高于普通PCR约100倍。将该方法应用于DHBV感染雏鸭的动力学研究,结果表明,DHBV感染雏鸭11 d时病毒含量达到高峰,随后逐渐下降。该方法的建立不仅为临床提供一种对DHBV的快速准确的检测方法,也为药物有效评价时间点的选择提供了理论依据。In this study,a SYBR Green fl uorescence quantifi cation PCR method was developed for the detection of Duck hepatitis B virus(DHBV).The specifi c primers were designed according to the C gene sequence of Duck hepatitis B virus.This method showed a good linear relationship when the template was among in 7.8×10^(2)-7.8×10^(8) copies/μL,and the correlation coeffi cient(R^(2))was 0.999.Total 95 clinical samples from different regions of Shandong were tested and the positive rate was 97.9%and its sensitivity was about 100 times higher than that of conventional PCR.The method was also applied to the kinetic study of DHBV infected ducklings.The results showed that the virus content reached a peak when DHBV infected ducklings on the 11th day and then gradually decreased.The development of this method not only facilitated a fast and accurate detection for DHBV but also provided a theoretical basis for the selection of effective drug evaluation.

关 键 词：鸭乙型肝炎病毒 荧光定量PCR 检测 感染动力学 

分 类 号：S852.659.6[农业科学—基础兽医学]