出口家禽新城疫病毒PCR和ELISA联合检测方法的建立

Establishment of PCR and ELISA combination test for Newcastle disease virus of export poultry

针对出口家禽新城疫泄殖腔棉拭样品病毒检测的需要，建立了IK2R和ELISA联合检测方法。结果表明：检测时间较常规缩短7～14d；通过被检样品鸡胚传代，提高了阳性检测率；特异性强，只与不同类型新城疫病毒反应，不与其他常见禽类病毒反应；所检的3216个样本，符合率98．8％；用多重RT-IK2R和脑内接种试验两种方法鉴定的114份样品，符合率99．1％；与G朗Bank中部分新城疫病毒F基因比较，同源性81％～100％。

A combined method of ELISA and RT-PCR to detect Newcastle Disease Virus （NDV） of export poultry was successfully established. Results of laboratory tests indicated that compared with a traditional method, this method could reduce the time spent in the whole test by 7-14 days and improve the detectable rate of positive NDV through chick embryo passage of specimens. The coincidence rate in the test for 3 216 specimens was 98.8%. The method had a good specificity, reacting only to different types of NDV and not to other common avian viruses. The coincidence rate between multiplex RT-PCR and intracerebral inoculation test for 114 specimens was 99.1%. The homology of F gene between the NDV isolates and the NDV strains of the GenBank was 81%-100% .