深圳市2005年鼠间汉坦病毒感染的鼠情监测及病毒株SZ2083的分离鉴定

目的 了解深圳市啮齿动物自然感染汉坦病毒（HV）状况和病毒型别，为制订防制措施提供科学依据。方法 收集宿主动物资料，采用酶联免疫吸附试验和间接免疫荧光试验分别检测鼠血清特异性总抗体和IgG抗体，直接免疫荧光检测鼠肺汉坦病毒抗原。将阳性鼠肺标本接种长爪沙鼠分离HV，阳性鼠肺标本提取病毒RNA，应用型特异性引物进行逆转录-巢式PCR扩增及核苷酸序列测定从而进行分型鉴定。结果 共捕获鼠形动物472只，其中以褐家鼠为优势鼠种，带病毒率为9．96％。鼠血标本总抗体阳性76例，IgG阳性56例。成功分离到1株HV，命名为SZ2083。经逆转录一巢式PCR扩增并进行序列测定显示为汉城（SEO）型。序列比较分析发现SZ2083核苷酸序列与L99同源性为97％，而与HTN76—118株的同源性仅为76％。结论 深圳市存在着以家鼠型为主的肾综合征出血热自然疫源地。

Surveillance on natural infection of rodents with Hantavirus in Shenzhen city and identification of a Hantavirus strain SZ2083

Objective For clarifying the situation of the natural infection of rodents having hemorrhagic fever with renal syndrome (HFRS) virus and to type Hantavirus (HV) using molecular technique in Shenzhen city in 2005,and offering guidance for prevention and control of HFRS.Methods Data on the host animals was collected from the city of Shenzhen.ELISA and indirect immunofluorscent antibody(IFA) test were applied to the specific antibodies against HV in the sera of captured rats.Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues of the HV infected rats were inoculated into Meriones unguiculata to isolate HV.The whole viral RNA was extracted from the lung tissues of the HV infected rats and amplified the partial M fragments with RT-nested-PCR,using the HV genotype specific primers.The amplified genes were then sequenced,and subjected to genotyping and homology analysis.Results 472 rodents were captured from Shenzhen in 2005.Surveillance on rats demonstrated 9.96% rats carrying HV (with a density of 8.25% ) and the main host was Rattus norvegicus.In the blood samples of rats,anti-HV IgG antibodies were detectable in 56 cases by IFA,and proved to be positive in 76 cases by ELISA.We successfully isolated a HV strain designated as SZ2083 from Rattus norvegicus for the first time in Shenzhen and was identified to SEO type by RT-nested-PCR. Compared with the coding region of the M gene of HV L99 virus strain,the homologies of nucleotide among them were 97%,but the homology was 76% of the SZ2083 with HTN 76-118 virus strain. Conclusion Results showed the existence of natural epidemic areas of HFRS in Shenzhen city.Based on the results of sequencing,it is possible that the Seoul strain of HV might be the predominant serotype of virus harbored.