数字农科院2.0

同时检测CSFV、PRRSV的二重RT-PCR方法的建立及其在临床样品检测中的初步应用

文献类型： 中文期刊

作者： 岳丰雄;崔尚金;冉多良;张超范;周盛华;戚亭

作者机构：

关键词： 多重PCR;RT-PCR;PRRSV;CSFV

期刊名称：畜禽业

ISSN： 1008-0414

年卷期： 2008 年 03 期

页码： 32-35

摘要： 建立了1种一步法RT-PCR用于同时检测典型猪瘟病毒(CSFV)和猪繁殖呼吸道综合征病毒(PRRSV)的多重PCR诊断方法(Multiplex PCR,mPCR)。根据Genbank上公布的CSFV、PRRSV基因组全序列及部分序列,借助DNAStar和Oligo6.0基因序列和引物设计软件,设计2对特异性引物分别用于扩增CFSV、PRRSV病毒777bp和434bp的目的片断。该mPCR由RT反应和PCR反应构成。将病毒核酸连接到PMD18-T载体,提取质粒后,以10倍进行倍比稀释,取每个稀释度的病毒核酸进行mPCR反应,对CSFV、PRRSV的最低检测量分别为8.5pg、8.3×10-2pg。以PCV2、PCV1、PPV、PRV、SIV、E.coil和双蒸水为模板进行mPCR反应,扩增结果均为阴性,表明该mPCR具有较好的特异性。对临床样品的检测表明,本研究建立的mPCR诊断方法能够对CSFV和PRRSV进行快速的诊断,同时对CSFV、PRRSV在猪群中的流行病学调查也具有十分重要的意义。

分类号： S854.43

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