深圳市HFRS疫源地鼠携带汉坦病毒与人感染汉坦病毒的基因特征对应性研究

摘要

目的 了解深圳市肾综合征出血热(HFRS)疫源地鼠携带汉坦病毒与人感染汉坦病毒二者的病原学特征及联系,为HFRS的防控提供科学依据.方法 收集疑似HFRS病人急性期血清标本,并在相同区域开展笼日法捕鼠,无菌取鼠肺.分别采用ELISA和直接免疫荧光法筛选阳性标本,选择代表性血清标本以及鼠肺标本进行逆转录-套式聚合酶链反应(RT-nested PCR)扩增部分M和S片段及核苷酸序列测定,与国内外的汉坦病毒毒株一起进行同源性比对和系统进化树分析.结果 472份鼠肺经直接免疫荧光检测共获得阳性鼠肺47份,带毒率为9.96%.对60份IgM抗体阳性的血液标本进行RT-nested PCR扩增,检出12份阳性,阳性率为20%.从代表性的8份鼠肺和8份病人血清中提取的标本二者之间M片段核苷酸序列的同源性高达95%以上,S片段之间的同源性高达96.5%以上.其推导的氨基酸序列同源性分别为98.0%～100%、98.4%～100%.深圳市HFRS疫源地鼠携带的汉坦病毒和人感染汉坦病毒均为汉城型S2亚型.结论 深圳市流行的汉坦病毒为SEO型S2亚型.无论是同一地区的鼠和人之间,还是不同地区的鼠和人之间,汉坦病毒都是具有较高的同源性.%Objective To compare and contrast the gene characteristic correspondence of hantaviruses(HV) carried by rats in natural epidemic areas of hemorrhagic fever with renal syndrome(HFRS) and infected among HFRS patients in Shenzhen,provide a reasonable scientific basis for controlling of HFRS.Methods We collected the patients serum specimens in acute stage and lung tissues samples of rats.ElISA and direct immunofluorescence were applied to screen the positive samples,respectively.The partial G2 fragments of M segment and S segment were amplified from the representative patients' serum positivesamples and lung tissues positive samples in different areas with reverse transcription-nested-polymerase chain reaction(RT-nested PCR) by the hantaviruses genotype specific primers.The amplified genes were then sequenced,and subjected to genotyping and homology analysis with other known hantaviruses.Results Four hundred and seventy-two rats were trapped in the main epidemic areas,and Hantavirus specific-antigens in lung tissues samples were identified in 47 out of the 472 by direct immunofluorescence.Twelve partial M and S segments were amplified from 60 patients serum specimens positive with specific IgM antibodies against hantavirus with ELISA by RT-nested PCR.The homology of M and S genome segments among 16 strains of Hantaviruses showed more than 95% and 96.5% at nucleotide level,respectively.And the deduced amino acid sequences homology was 98.0% -100% and 98.4%-100%,respectively.The genotype of hantavirus carried by rats and infected among patients were identified to the same subtype-SEO S2.Conclusion The genotype of Hantavirus carried by rats and infected among patients in Shenzhen all belongs to S2 SEOV.The nucleotide homology of SEO type of Hantavirus in the same or nearby areas is higher and the viruses are highly conserved.