关键词: 汉坦病毒, 细胞培养, 复苏, 鉴定

Abstract:

Objective To understand the distribution characteristics of Hantavirus (HV) genotype in Hebei province, the HV strains isolated from rodents in 2012 and recovered from formerly saved were typed. Methods Lung samples were screened by immunoinfluscent assay (IFA). Antigen-positive lung samples were processed and inoculated onto cultures of confluent Vero-E6 cells. Formerly saved strains were recovered. Total RNA were extracted for genotype and identification. Results Four lung tissues were homogenized and inoculated onto confluent Vero-E6 cells. After the second-third passage, the characteristic HV antigen particle could be seen by IFA. At last, all four strains were isolated successfully. The formerly saved strains were used to infect fresh Vero-E6 cell cultures. After the first-second passage, the characteristic HV antigen particle can be seen by IFA. At last, all six strains were recovered successfully. All ten strains were typed by PCR and they belonged to SEO type. Conclusion SEO type was the predominant HV genotype in Hebei province. SEOV didn't produce cytopathic effects, but the virus can proliferate in the cell. HV can be saved for at least more than 20 years at -80 ℃. In China, Microtus fortis was an alternative host for SEOV.

Key words: Hantavirus, Cell culture, Recovery, Identification