轮状病毒基因重配株血清型鉴定方法的研究

Serotype Canidate Reassortant Rotavirus Vaccine Strains by a Novel RT-PCR Dot-Blot Hybridizatation Method

Ａ组轮状病毒中根据基因９的不同目前至少已发现有１３个不同Ｇ血清型，其中能引起人类致病的有Ｇ１－Ｇ４，Ｇ８，Ｇ９和Ｇ１２型。建立可靠的血清型鉴定技术对于轮状病毒疫苗的研制和分子流行病学的研究具有重要意义。本文首次报导了一种鉴定轮状病毒Ｇ血清型的新方法，利用已知有关轮状病毒ＶＰ７基因的序列资料，设计合成了一套鉴定轮状病毒Ｇ血清型的寡核苷酸探针，利用地高辛标记上述探针。待检品经反转录ＰＣＲ扩增后与上述一套寡核苷酸探针分别进行杂交得以确定其血清型。这一方法与目前常用的套式ＰＣＲ方法相比更适合于大量样品的操作而且结果可靠。用这一方法对本实验室组建的四株基因重配疫苗株进行实验，其结果与套式ＰＣＲ方法完全一致。

There are at least thirteen genetically distinct G serotypes in group A rotavirus on the basis of seological evidencce and comparative nuceotide sequencing and the predicaed animo acid sequence of VP7 gene.To identify each VP7 serotype of the reassortant rotavirus vaccine candidate strains which we selected before,we developed a novel method.Four oligonucleotide probes were designed based on the sequence information available for VP7 serotypes.After the ds RNA of rotavirus samples were reverse transcribed into ds DNA,a hybridization test were carred out by using the four oligonucleotice probes respectively strain.Our results suggest that this method will be useful in epidemiloogic research and vaccine-related interest.