北方果树主要病毒聚合酶链式反应检测试剂盒的研制

Development of Polymerase Chain Reaction Kit for Northern Fruit Tree’s Main Virus

为使检测ACLSV、ASGV、ASPV、PNRSV和PDV5种果树主要病毒达到最优化检测。本研究以GF305、弗吉尼亚小苹果带毒(阳性)和非带毒(阴性)为试材,提取总RNA,根据ACLSV、ASGV、ASPV、PNRSV和PDV这5种病毒的外壳蛋白基因序列设计引物,利用逆转录-聚合酶链式反应(RT-PCR)技术进行检测,最终研制出上述5种果树主要病毒PCR检测试剂盒。利用本试剂盒(该试剂盒已获得国家发明专利授权(ZL200710010565.8)检测果园中存在的病毒,明确了果园中这5种病毒的感染率,实现了快速、灵敏、成本低、特异性强的检测目的。

In order to detect the ACLSV,ASGV,ASPV,PNRSV and PDV and achieve optimal detection.Total RNA was extracted from GF305 and Virginia crab.Primers were designed according to the coat protein gene sequences of ACLSV,ASGV,ASPV,PNRSV and PDV,and the reverse transcription-polymerase chain reaction technique was used for detection.Ultimately,PCR detection kits for 5 main viruses in fruit trees were developed.The virus in orchard was detected by this kit(The kit has obtained the national invention patent authorization(ZL200710010565.8),and the infection rate of these 5 viruses in orchards was determined,and the rapid,sensitive,low cost and high specificity detection were achieved.