一种简单高效扩增人杯状病毒ORF2区RT-PCR方法的建立及其意义

人杯状病毒(human calicivirus,HuCV)属于杯状病毒科(Caliciviridae),是单股正链RNA病毒,长约7·5 kb,其3′末端有poly(A)结构。它可分为两个属:诺如病毒(Norovirus)和札如病毒(Sapovirus)[1],根据病毒抗原性和核苷酸序列的多样性,目前将诺如病毒和札如病毒分别划分为三个遗传组(group),每一遗传组依据RNA多聚酶及衣壳蛋白区域序列的差异,可进一步划分为不同群或基因型(cluster or genotype)。病毒基因组包括3个开放读码框(open reading frame,ORF),5′端和3′端各有一个小的非编码区。ORF1编码非结构蛋白的前体聚蛋白,其中包括RNA…

Establishment and the Significance of a Simple and High Performance RT-PCR Method to Amplify ORF2 of Human Calicivirus

The capsid protein encoded by ORF2 of human calicivirus(HuCV) expressed in the baculovirus expression system can self-assembled into virus-like particles(VLPs),which would be important for the development of improving diagnostic methods.The RT-PCR procedures routinely used to amplify the RNA polymerase region have usually been inefficient for amplification of ORF2,which would bring obstacle for the development of new diagnostic reagents.Through optimization of RT-PCR conditions,we established a simple and high performance RT-PCR method to amplify ORF2,and by which demonstrated a corresponding relationship between the genotype and the antigenic type.It provides conditions for developing diagnostic reagent which accommodates to diagnosis of HuCVs prevailing in China,and also provides the basis for studying the genetic variance and the pathogenesis of HuCV.