1型星状病毒上海分离株基因组序列测定及分析

目的 了解上海地区腹泻患儿感染的星状病毒基因组分子结构特点和基因型.方法 用RT-PCR方法分段扩增星状病毒基因组,克隆于pMD18-T载体上,测定序列并拼接,用MEGA、DNAStar软件对所得序列进行分析.结果 本次分离株基因组全长6807 bp,命名为HAstV-SH,提交到GenBank上的序列号为FJ375759,有3个开放阅读框架,非结构基因ORF1a、ORF1b共长4290 bp,位于基因组83～4372 nt之间;编码结构蛋白的ORF2基因全长2364 bp,位于基因组4764～6727 nt之间.与GenBank中星状病毒ORF2基因组序列同源性比较发现,HAstV-SH与1型星状病毒核苷酸同源性最高(97%),与其他基因型的同源性为63%～70%.结论 上海地区儿童感染的星状病毒HAstV-SH属于1型星状病毒,与日本分离株AB009985亲缘关系较近,氨基酸同源性为97.5%.

The genomic sequence analysis of human astrovirus in Shanghai

Objective To study the genomic molecular organization and genotype of human astro-virus infected infants in Shanghai of China. Methods Based on the published genomic sequence of HAstV (GenBank), the whole genome of one isolate human astrovirus was sequenced by specific primers. The PCR-products were cloned to pMD18-T vector and sequenced, phylogenetic tree was constructed by the Neighbor-Joining method with MEGA 4 software. Results The genome of HAstV-SH is 6807 hp, contains three ORFs: ORF1a and 1b encode the non-structural protein (from 83 nt to 4372 nt), ORF2 encodes the structural protein (from 4364 nt to 6727 nt). Compared with the ORF2 gene of those eight astrovirus sero-types in GenBank, revealed that the highest homology is with genotype 1 (97%). Homology with other gen-otypes ranged between 63% and 70%. Conclusion HAstV-SH belonged to genotype 1 and closely clus-tered with a strain of Japan (AB009985).