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鸡传染性喉气管炎病毒实时荧光PCR检测方法的建立与应用

19067 2020-03-12 14:12:05 王楷宬,王素春,孙亚伟,等 中国动物检疫
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为探索适合临床高通量鸡传染性喉气管炎检测的方法,针对鸡传染性喉气管炎病毒gB基因序列保守区域,筛选1对特异性引物和1条特异性探针,建立了实时荧光PCR检测方法,并对该方法进行特异性、敏感性评估和临床应用检测。结果显示:筛选出的引物和探针与其他禽类常见病毒无交叉反应,检测下限达到100拷贝/反应;对来自13个场的480份临床样本进行检测,检测出17个阳性样品,与常规PCR检测结果符合率为100%。结果表明,此方法特异性强,灵敏度高、耗时少,可用于鸡传染性喉气管炎的快速检测。

Establishment and Application of a Real-time PCR for Detecting Infectious Laryngotracheitis Virus

In order to develop an appropriate method for high-throughput detection of avian infectious laryngotracheitis(AILT),a real-time PCR assay was established with a pair of specific primers and a probe based on the conservative zone of gB gene sequence of AILT,then its specificity and sensitivity were evaluated,and its clinical application was tested. The results showed that the selected primers and probe failed to cross-react with other common poultry viruses,with the detection limit of 100 copies/reaction. 17 out of 480 clinical samples collected from 13 farms were positive. The results completely conformed to the test result by common PCR assay. As a conclusion,AILT could be rapidly detected by the established method with high specificity,high sensitivity and less time-consuming.

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