首个仁果类果树主要病毒RT-PCR检测试剂盒的研制

Development of the First Virus RT-PCR Detection Kit for Pome Fruits

以GF305、弗吉尼亚小苹果为带毒(阳性)和非带毒(阴性)试材,利用作者已建立的总RNA提取方法提取试材中的总RNA,根据三种病毒(ACLSV,ASGV和ASPV)外壳蛋白基因序列设计寡核苷酸引物,采用RT-PCR技术研制三种病毒检测试剂盒,达到最优化检测。并应用该试剂盒对果园中存在的病毒进行了检测。该试剂盒检测病毒速度快(6~7 h)、灵敏度高(较ELISA法高8 000倍)、专一性强、检测成本较低,是一种有推广价值的仁果类果树主要病毒检测方法。

GF305 and Virginia small apples were taken as toxic（positive） and non toxic（negative） samples,and the total RNA of the samples were extracted by the established total RNA extraction method.Oligonucleotide primers were designed according to the coat protein gene sequences of three viruses（ACLSV,ASGV and ASPV）.RT-PCR technology was used to develop three kinds of virus detection kit to achieve the optimal detection,which were applied to detect the virus in orchard.The results showed that this virus detection kit had faster speed（6~7 h）and higher sensitivity（8 000 times higher than ELISA）,stronger specificity,and lower detection cost,which could be the main virus detection method of pome fruit trees with promotion value.