多方法组合调查新疆阿勒泰地区6种常见鼠传致病菌流行状况  

Prevalence of 6 common rodent-borne pathogens identified using multiple methods in Xinjiang Altay prefecture,China

作　　者：李瑞晓 陈燕芳 穆龙 徐爱玲 刘蓬勃 王君[2] 马伟 栗冬梅[2] LI Ruixiao;CHEN Yanfang;MU Long;XU Ailing;LIU Pengbo;WANG Jun;MA Wei;LI Dongmei(School of Public Health,Cheeloo College of Medicine,Shandong University,Jinan 250012,Shandong,China;State Key Laboratory of Infectious Disease Prevention and Control,Department of Vector Biology and Control,National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Institute for Agricultural Sciences of 10th Division of Xinjiang Production and Construction Corps,Beitun 836099,Xinjiang,China)

机构地区：[1]山东大学齐鲁医学院公共卫生学院,山东济南250012 [2]中国疾病预防控制中心传染病预防控制所媒介生物控制室传染病预防控制国家重点实验室,北京102206 [3]新疆生产建设兵团第十师农业科学研究所,新疆北屯836099

出　　处：《微生物学通报》2023年第9期4109-4124,共16页Microbiology China

基　　金：传染病预防控制国家重点实验室面上项目(2022SKLID202);国家科技重大专项(2017ZX10303404);国家自然科学基金面上项目(31970005)。

摘　　要：【背景】鼠传疾病是对人类危害较大的一种人兽共患病,全球化使得鼠传疾病流行区域不断扩大,出现了多种新发鼠传疾病的发生及旧传染病的复燃。【目的】调查新疆阿勒泰地区常见的鼠传致病菌在啮齿动物中的流行状况,为当地自然疫源性疾病防控提供科学依据。【方法】采用夹夜法捕获啮齿动物,无菌收集其脾脏和肾脏组织,提取基因组DNA。应用TaqMan探针法的荧光定量聚合酶链式反应(quantitative polymerase chain reaction,qPCR)检测巴尔通体(Bartonella spp.)、问号钩端螺旋体(Leptospira interrogans)、恙虫病东方体(Orientia tsutsugamushi)、莫氏立克次体(Rickettsia mooseri)、嗜吞噬细胞无形体(Anaplasma phagocytophilum)和土拉弗朗西斯菌(Francisella tularensis)6种常见的鼠传致病菌。采用16S rRNA基因的通用引物进行常规PCR扩增后,应用Illumina测序和Nanopore测序进一步检测致病菌,同时对脾脏组织进行巴尔通体体外分离培养。比较qPCR、16S rRNA基因测序和分离培养的结果。【结果】共捕获啮齿动物8种66只,其中,乌拉尔姬鼠(Apodemus uralensis)31只,占捕获总数的46.97%,其余为褐家鼠(Rattus norvegicus)、小家鼠(Mus musculus)等。qPCR检测常见的鼠传致病菌感染率为巴尔通体31.80%(21/66)、问号钩端螺旋体1.50%(1/66)、恙虫病东方体1.50%(2/66)、莫氏立克次体3.00%(1/66)和土拉弗朗西斯菌13.60%(9/66),未检出嗜吞噬细胞无形体。16S rRNA基因Illumina测序分析通过质控的23份样品检测出致病菌,以巴尔通体为主;16S rRNA基因Nanopore测序分析通过质控的11份样品检测出巴尔通体,其他5种常见的鼠传致病菌未检出。有11份脾脏组织分离培养出巴尔通体,感染率为16.67%(11/66)。【结论】新疆阿勒泰地区的啮齿动物可携带巴尔通体等多种鼠传致病菌,应关注并加强该地区相关传染病的防治工作。qPCR、细菌分离培养、16S rRNA基因测序3种检测方�[Background]Rodent-borne diseases are a class of zoonoses harmful to humans.The epidemic areas of rodent-borne diseases keep expanding with the progress in globalization,and a variety of new rodent-borne diseases emerge while the old infectious diseases reoccur.[Objective]To investigate the prevalence of common rodent-borne pathogens in rodents in Altay prefecture of Xinjiang,and to provide a scientific basis for the prevention and control of local natural focus diseases.[Methods]The spleen and kidney tissue samples were collected aseptically from the rodents captured by night trapping method,and the genome DNA was extracted.Six common rodent-borne pathogens including Bartonella spp.,Leptospira interrogans,Orientia tsutsugamushi,Rickettsia mooseri,Anaplasma phagocytophilum,and Francisella tularensis were detected by fluorescence quantitative polymerase chain reaction(qPCR)with TaqMan probe.Illumina sequencing and Nanopore sequencing after routine PCR amplification with universal primers for the 16S rRNA gene were employed to further detect the pathogens.Meanwhile,the spleen tissue was used for the isolation and culture of Bartonella in vitro.The results of qPCR,16S rRNA gene sequencing,and bacterial isolation and culture were compared.[Results]A total of 66 rodents of 8 species were captured,of which 31(46.97%)rodents were Apodemus uralensis,and the rest were Rattus norvegicus,Mus musculus,etc.The infection rates of common rodent-borne pathogens detected by qPCR were as follows:Bartonella 31.80%(21/66),L.interrogans 1.50%(1/66),O.tsutsugamushi 1.50%(2/66),R.mooseri 3.00%(1/66),and F.tularensis 13.60%(9/66).A.phagocytophilum was not detected.The Illumina sequencing of 16S rRNA gene detected pathogens(mainly Bartonella)in the 23 samples passing the quality control.The Nanopore sequencing of 16S rRNA gene detected Bartonella in 11 samples passing the quality control,and did not detect the other 5 common rodent-borne pathogens.Bartonella was isolated from 11 spleen tissue samples,with the infection rate of 16.67%(11/

关 键 词：啮齿动物 鼠传致病菌 定量聚合酶链式反应 16S rRNA基因测序 Illumina测序 Nanopore测序 

分 类 号：S852.6[农业科学—基础兽医学] R378[农业科学—兽医学]