深圳市啮齿动物感染汉坦病毒的基因特征研究

罗瑶; 李玥; 黄亚兰; 张晓敏; 熊玲红; 张仁利; 阳帆

Abstract

罗瑶,李玥,黄亚兰,张晓敏,熊玲红,张仁利,阳帆.深圳市啮齿动物感染汉坦病毒的基因特征研究[J].Chinese journal of Epidemiology,2022,43(11):1804-1810

深圳市啮齿动物感染汉坦病毒的基因特征研究

Genetic characteristics of hantavirus detected in rodents in Shenzhen

Received:June 29, 2022

DOI：10.3760/cma.j.cn112338-20220629-00576

KeyWord: 汉坦病毒汉城病毒基因分型序列分析

English Key Word: Hantavirus Seoul virus Genetic type Sequence analysis

FundProject:

Author Name	Affiliation	E-mail
Luo Yao	School of Public Health, Southern Medical University, Guangzhou 510515, China Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China
Li Yue	Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China
Huang Yalan	Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China
Zhang Xiaomin	Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China
Xiong Linghong	Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China
Zhang Renli	School of Public Health, Southern Medical University, Guangzhou 510515, China Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China
Yang Fan	School of Public Health, Southern Medical University, Guangzhou 510515, China Institute of Pathogenic Biology, Shenzhen Center for Disease Prevention and Control, Shenzhen 518055, China	sailing_728@163.com

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Abstract:

目的研究深圳市啮齿动物所携带的汉坦病毒分子流行病学特征，分析汉坦病毒基因型别。方法采用笼日法布点捕鼠，收集鼠肺标本后研磨提取RNA。运用实时荧光PCR方法进行分型检测，进一步采用反转录-巢式PCR扩增部分M片段G2区和S片段核苷酸序列，并进行同源性和系统进化树分析。结果共捕获200只鼠类动物，包含褐家鼠189只、黄胸鼠9只和小家鼠2只。汉坦病毒检出率为21.0%（42/200），均为汉城病毒，其中宝安区鼠肺检出率最高，达45.7%（χ²=25.60，P<0.05）。从阳性鼠肺标本中分别扩增出25条汉坦病毒M片段G2区和S片段序列，核苷酸同源性分别为95.3%~100.0%和97.6%~100.0%，与来自广州市的参考序列核苷酸相似性较高。系统进化树显示此次研究深圳市鼠类动物所携带的汉坦病毒均为汉城病毒的S2亚型。氨基酸突变位点分析发现，在S基因编码的核衣壳蛋白中存在1个位点突变，为BA-111第973位由丙氨酸（Ala）变为苏氨酸（Thr）。结论深圳市鼠类动物所携带的汉坦病毒为汉城病毒的S2亚型，与深圳市历年以及周边省市汉坦病毒代表病毒毒株相比变异不大。

English Abstract:

Objective To study the molecular epidemiological characteristics and genotypes of hantavirus carried by rodents in Shenzhen. Methods Rodents were captured, and their lung samples were collected and grinded for RNA extraction. The hantavirus positive samples were classified by real-time fluorescence PCR. Rat lung nucleic acid samples were selected to amplify the nucleotide sequences of partial M fragments (G2 segment) and S fragments by reverse transcription-nested polymerase chain reaction (RT-nested PCR). The PCR products were then sequenced and homology and phylogenetic tree analyses were conducted. Results A total of 200 rodents were captured, including 189 Rattus norvegicus, 9 Rattus flavipectus and 2 Mus musculus. The positive rate of hantavirus was 21.0% (42/200), all of the isolates were seoul virus (SEOV) strains. The positive rate of hantavirus in Bao'an district was highest (45.7%), and the difference in detection rate among districts were significant (χ²=25.60,P<0.05). A total of 25 G2 segment sequences and S fragment sequences of SEOV were obtained by virus gene sequencing, and their nucleotide homology was 95.3%-100.0% and 97.6%-100.0%, respectively. Compared with other reference sequences of S2 subtype, the nucleotide homology between the sample sequence and the reference sequence from Guangzhou was high. Analysis on nucleotide homology and phylogenetic tree showed that hantavirus carried by the rodents captured in Shenzhen belonged to SEOV S2 subtype. Analysis on amino acid variation sites revealed that there was a variation in the nucleocapsid protein encoded by S gene from Alanine to Threonine at the 973 position of BA-111.Conclusion Hantavirus carried by rodents in Shenzhen belongs to S2 subtype of Seoul virus, which have little variation compared with the hantavirus strains obtained in other years in Shenzhen and surrounding provinces.

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