摘要
对同样以引起鸭产蛋下降为主要特征的鸭流感病毒(AIV)分离株和鸭新城疫病毒(NDV)YH99V分离株,通过设计特异性引物和优化体系反应条件,建立了一种早期快速鉴别诊断鸭AIV和鸭NDV的单项RT-PCR方法和多重RT-PCR方法,AIV的扩增片段大小为483bp,NDV的扩增片段为310bp,电泳快速,易于区分。敏感性试验表明,该方法比HA敏感100倍以上;对AIV、YH99V、IBV、IBDV及正常鸡胚尿囊液的混合液检测显示,多重RT-PCR法具有很强的特异性。对鸭病毒性产蛋下降征侯群送检病料的检测证明,该方法能有效鉴别AIV和/或NDV单独感染或混合感染,阳性检出率明显高于HA方法。
In order to differentiate the isolate of AIV from isolate YH99V of NDV, both of which were isolated from duck and generally cause egg to sharp-drop in laying ducks, a rapid diagnostic techniques of multiple RT-PCR was established by designing the specific primer and optimizing the reaction condition. The DNA segments amplified were 483 bp in length for AIV and 310 bp in length for NDV, which could be conveniently differentiated by the agarose gel electrophoresis. Sensitivity test results indicated that as little as 1/100 of HA titer could be detected by the multiple RT-PCR. Detection of mixed allantoic fluids containing AIV, YH99V, IBV, IBDV and negative allantoic fluids showed that the multiple RT-PCR possessed a high specificity. Detection of field samples from viral diseased ducks with egg drop also confirmed that the multiple RT-PCR could identify the single infection of AIV, NDV and combined infection of the both. And the positive detection ratio of the multiple RT-PCR was significantly higher than that of HA method.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2007年第3期296-299,共4页
Chinese Journal of Veterinary Science
基金
浙江省自然科学基金资助项目(302077)
浙江省科技厅项目(2004C32039)
浙江出入境检验检疫局项目(ZK200613)
