摘要
为满足新城疫病毒(NDV)快速检测的需求,通过对我国流行的6种基因型NDV基因组序列进行比对,针对P基因保守区域设计特异性引物和探针,建立了NDV通用荧光RT-PCR检测方法,并评估了方法的特异性、敏感性、重复性和诊断性能。结果显示:该方法可检测目前国内流行的6种基因型NDV,与其他常见禽病病毒无交叉反应,灵敏度比常规RT-PCR高10倍,组内和组间变异系数均低于1.5%。利用该方法检测1044份临床样品,绘制ROC曲线,其线下面积为0.9718,诊断的敏感性和特异性分别为92.62%和98.99%,与病毒分离鉴定结果的符合率为98.08%。结果表明,本研究建立的方法特异性强、敏感性高、重复性好、准确性高,可用于NDV的快速、通用检测。
In order to rapidly detect Newcastle disease virus(NDV),a universal fluorescent RT-PCR assay was established based on specific primers and probes designed targeting the conserved region of P gene that was selected through comparing the genomic sequences of 6 genotypes of strains prevalent in China,and assessed for its specificity,sensitivity,repeatability and diagnostic performance.The results showed that the method could be used to detect all the 6 genotypes of NDV,but failed to crossly react with other common avian disease viruses,its sensitivity was 10 times higher than that of conventional RT-PCR,and the coefficients of variation(CV)for both intra-and inter-assay repeatability were less than 1.5%.1044 clinical samples were tested by the method,and the area under the ROC curve was 0.9718,the diagnostic sensitivity and specificity were 92.62%and 98.99%,respectively,and the coincidence rate between the new method and virus isolation and identification method was 98.08%.In conclusion,the established method,with its high specificity,sensitivity,reproducibility and accuracy,could be used for rapid and universal detection of NDV.
作者
王静静
克军宏
王海鸣
郭通
于晓慧
刘华雷
Wang Jingjing;Ke Junhong;Wang Haiming;Guo Tong;Yu Xiaohui;Liu Hualei(China Animal Health and Epidemiology Center,Qingdao 266032,Shandong,China;Key Laboratory of Animal Biosafety Risk Warning,Prevention and Control(Southern China),Ministry of Agriculture and Rural Affairs,Qingdao 266032,Shandong,China;Ningxia University,Yinchuan 750021,Ningxia,China;Qingdao Agricultural University,Qingdao 266109,Shandong,China)
出处
《中国动物检疫》
2025年第11期83-88,96,共7页
China Animal Health Inspection
基金
国家重点研发计划项目(2022YFD1800600)。
