鼠类不同组织巴尔通体的分离培养及其遗传多态性分析  

Isolaltion of Bartonella in different tissues of rodents and analysis of its genetic diversity

作　　者：刘海鑫 王加熊 刘维俊 刘菁 徐国英 郑奎城[1,2] 肖方震[1,2] LIU Hai-xin;WANG Jia-xiong;LIU Wei-jun;LIU Jing;XU Guo-ying;ZHENG Kui-cheng;XIAO Fang-zhen(Fujian Provincial Key Laboratory of Zoonosis Research,Fujian Center for Disease Control and Prevention,Fuzhou 350001,China;School of Public Health,Fujian Medical University)

机构地区：[1]福建省疾病预防控制中心福建省人兽共患病重点实验室,福建福州350001 [2]福建医科大学公共卫生学院

出　　处：《中国病原生物学杂志》2022年第3期293-297,302,共6页Journal of Pathogen Biology

基　　金：国家科技重大专项课题项目(No.2017ZX10103008);福建省卫生健康中青年骨干人才培养项目(No.2019-ZQN-27);福建省科技创新平台建设项目(No.2019Y2001)。

摘　　要：目的分析鼠类不同组织巴尔通体的分离率及培养时间,了解福建省巴尔通体分离株的遗传多态性。方法采用笼日法捕获家栖鼠和野栖鼠,现场鉴定鼠种,无菌条件下采集血液、脾、肝、肾和肺等组织进行巴尔通体分离培养,挑选疑似菌落进行分子生物学鉴定,分析不同组织巴尔通体分离率及培养时间;PCR扩增巴尔通体的gltA基因,测序后进行系统发育分析和遗传多态性分析。结果在78只鼠中,巴尔通体感染率为56.41%。血、脾、肝、肾和肺组织分离率分别为50.00%,44.87%,44.87%、48.72%和26.92%,不同组织巴尔通体分离率不同(χ^(2)=11.042,P<0.05),肺组织分离率较低。肾和血液分离率结果具有一致性(Kappa=0.821,P<0.01)。阳性菌落最早长出时间为2 d,多数在1周内生长,不同组织巴尔通体分离时间差异有统计学意义(χ^(2)=14.169,P<0.01),肝组织中巴尔通体分离培养所需时间最长。不同巴尔通体的菌落长出时间的差异有统计学意义(χ^(2)=21.249,P<0.01),昆州巴尔通体的菌落长出所需时间最长。遗传多样性分析显示,福建巴尔通体分离株单倍型多样性(Hd)和核苷酸多样性指数(π)的值分别为Hd=0.747±0.054,π=0.04166±0.00611。结论巴尔通体具有多组织嗜性,血液和肾脏可作为巴尔通体分离培养的首选。不同组织和不同种型分离巴尔通体,其菌落长出时间不同。福建巴尔通体分离株遗传多样性丰富。Objective To analyse the ioslation rate and ioslation time of Bartonella in different tissues of rodents and understand the genetic diversity of Bartonella species in Fujian Province.Methods Rodents were captured using live-capture traps.Traps were pre-baited with corns and set before sundown and then collected before sunrise.After the trapped rodent were morphologically examined,The blood,spleen,liver,kidney and lung were collected under sterile conditions for isolation and culture of Bartonella.The like-Bartonella colonies were selected for molecular identification.The isolation rate and culture time of Bartonella from different tissues were analyzed.The gltA gene fragment was amplified using PCR,After positive PCR products were sequenced,the phylogenetic tree was created and the genetic diversity of Bartonella was analyzed.Results A total of 78 rodents were captured,the prevalence of Bartonella in rodents was 56.41%.The isolation rates of blood,spleen,liver,kidney and lung tissues were 50.00%,44.87%,44.87%,48.72%and 26.92%,respectively.The isolation rate of Bartonella was different in different tissues of rodent(χ^(2)=11.042,P<0.05),and the isolation rate of lung tissue was low.The result of kidney and blood isolation rate were highly consistent(Kappa=0.821,P<0.01).The typically observed visible Bartonella colonies from primary isolations can be obtained as soon as 2 days,most of them grow within 1 week.The incubation time of colonies in different tissues(χ^(2)=14.169,P<0.01)and different Bartonella species(χ^(2)=21.249,P<0.01)was different,and the isolation of Bartonella in liver tissues required a longer time than other tissues,and the incubation time of B.queenslandensis was longest.Genetic diversity analysis showed that the values of haplotype diversity(Hd)was 0.747±0.054 and nucleotide diversity index(π)was 0.04166±0.00611.Conclusion Bartonella can invade various tissues.Blood and kidney can be the first choice for Bartonella isolation and culture.The incubation time of Bartonella colonies isolated

关 键 词：巴尔通体 鼠类 不同组织 分离时间 遗传多样性 

分 类 号：R37[医药卫生—病原生物学]