2021-2022年我国新疆维吾尔自治区家畜流行性乙型脑炎病毒检测  

Detection of Japanese encephalitis virus in livestock in Xinjiang Uygur Autonomous Region of China from 2021 to 2022

作　　者：李冰 高岩[2,3] 曹鑫宇 仇相书 秦爱建 张赫[3] 金宁一 LI Bing;GAO Yan;CAO Xinyu;QIU Xiangshu;QIN Aijian;ZHANG He;JIN Ningyi(Jiangsu Key Laboratory of Animal Preventive Medicine,College of Veterinary Medicine,Yangzhou University,Yangzhou,Jiangsu 225001,China;Agricultural College,Yanbian University,Yanji,Jilin 133002,China;Changchun Institute of Veterinary Medicine,Chinese Academy of Agricultural Sciences,Changchun 130117,China;Institute of Pathogenic Biology,Peking Union Medical College,Beijing 100006,China;College of Animal Science,Zhejiang University,Hangzhou 310030,China)

机构地区：[1]扬州大学兽医学院江苏省动物预防医学重点实验室,江苏扬州225001 [2]延边大学农学院,吉林延吉133002 [3]中国农业科学院长春兽医研究所,吉林长春130117 [4]北京协和医学院病原生物学研究所,北京100006 [5]浙江大学动物科学学院,浙江杭州310030

出　　处：《中国兽医学报》2024年第8期1713-1718,共6页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2022YFD1800100)。

摘　　要：基于建立的SYBR GreenⅠ实时荧光定量PCR检测方法,检测新疆部分地区家畜血清样品中流行性乙型脑炎病毒(Japanese encephalitis virus,JEV)的流行情况。利用PCR扩增JEV毒株的E基因片段,将其克隆到pEASY-Blunt载体,构建重组质粒,建立SYBR GreenⅠ实时荧光定量PCR检测方法,并验证其灵敏度、特异性和重复性。收集2021-2022年新疆哈密、阿勒泰、伊犁、阿克苏、喀什地区家畜血清样品,应用建立的检测方法进行检测并分析阳性率,以监测新疆地区JEV在家畜中的流行情况。结果显示,建立的检测方法呈良好的线性关系,检测区间在4.03×10^(2)~4.03×10^(9)拷贝/μL,相关系数为0.995,斜率为-3.431,灵敏度下限极值为4.03×10^(2)拷贝/μL。该方法用于检测寨卡病毒(Zika virus,ZIKV)、登革病毒(Dengue virus,DENV)未出现特异性扩增。重复性的组内变异系数为0.53%~1.27%,组间变异系数为0.48%~1.43%。使用本方法检测2021-2022年新疆地区家畜血清样品,总阳性率为3.11%,其中马、牛和羊中的阳性检出率分别为2.35%、5.26%和3.74%,鉴定为基因Ⅰ型JEV。结果表明,建立了基因Ⅰ型JEV的SYBR GreenⅠ实时荧光定量PCR检测方法,新疆地区马、牛、羊血清中均检测到JEV,总阳性率为3.11%。The SYBR Green Ⅰ real-time fluorescence quantitative PCR detection method was used to determine the prevalence of Japanese encephalitis virus(JEV) in mosquito vectors and cattle serum samples in Xinjiang.The E gene fragment of the JEV strain was amplified by PCR,cloned into a pEASY-Blunt vector,produced as a recombinant plasmid,and its sensitivity,specificity and reproducibility were verified.Between 2021 and 2022,serum samples were taken in the regions of Hami,Altay,Ili,Aksu,and Kashi in order to monitor the prevalence of JEV in livestock in Xinjiang.The positive rate was discovered and evaluated using the established detection method.The established detection method showed a good linear relationship,and the detection interval was 4.03×10^(2)-4.03×10^(9) copies/μL.The correlation coefficient was 0.995,the slope was-3.431,and the extreme value of the lower limit of sensitivity was 4.03×102 copies/μL.This method has no specific amplification for Zika virus(ZIKV)and Dengue virus(DENV).The intra group coefficient of variation of reproducibility was 0.53%-1.27%,and the inter group coefficient of variation was0.48%-1.43%.Using this method to detect serum samples from livestock in Xinjiang from 2021to 2022,the total positive rate was 3.28%,with positive detection rates in horses,cows,and sheep being 2.35%,6.77%,and 3.74% respectively,the virus was identified as Type I JEV.A SYBR Green Ⅰreal-time PCR method for the detection of genotype I JEV was established.JEV was detected in the serum of horses,cattle and sheep in Xinjiang,with a total positive rate of 3.11%.

关 键 词：流行性乙型脑炎病毒 流行病学 E基因 检测方法 

分 类 号：R512.32[医药卫生—内科学] S852.65[医药卫生—临床医学]