Molecular cloning and nucleotide sequence analysis of the S-segment in Hantavirus isolated in Zhejiang province

VernacularTitle:1株汉坦病毒的分离及其S基因片段的克隆和序列分析
Author: Fang XU ; Pingping YAO ; Hanping ZHU ; Ronghui XIE
Publication Type:Journal Article
Keywords: Hantavirus; isolation; S segment; sequence analysis
From: Chinese Journal of Zoonoses 2009;(7):665-668
CountryChina
Language:Chinese
Abstract: One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.