2018-2019年福建省人感染布鲁氏菌分离株种型鉴定及分子特征分析  

Species identification and analysis of the molecular characteristics of Brucella strains isolated from humans in Fujian,China in 2018-2019

作　　者：韩腾伟[1] 林代华[1] 陈志平 刘菁[1] 刘维俊[1] 肖方震[1] 邓艳琴[1,2] HAN Teng-wei;LIN Dai-hua;CHEN Zhi-ping;LIU Jing;LIU Wei-jun;XIAO Fang-zhen;DENG Yan-qin(Fujian Center for Disease Control and Prevention,Fujian Provincial Key Laboratory for Zoonosis Research,Fuzhou 350001,China;College of Public Health,Fujian Medical University,Fuzhou 350122,China)

机构地区：[1]福建省疾病预防控制中心,福建省人兽共患病研究重点实验室,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》2021年第9期1001-1007,共7页Journal of Pathogen Biology

基　　金：国家科技重大专项项目(No.2018ZX10734401-007);福建省自然科学基金项目(No.2016J01347);福建省科技创新平台建设项目(No.2019Y2001)。

摘　　要：目的探讨福建省人感染布鲁氏菌分离株主要流行株的种型和分子特征,为制定预防控制策略提供依据。方法将布鲁氏菌分离株转种培养并提取基因组DNA,采用传统生物学鉴定和BCSP31-PCR、AMOS-PCR、MLST及MLVA-16等方法进行布鲁氏菌分离株分子鉴定和分型,通过Bionumerics 6.6软件对其进行聚类分析。结果2018-2019年福建省人感染布鲁氏菌22株分离株分子检测结果与传统分型基本相符,为2个种(羊种和猪种)和2个生物型(羊3型和猪3型),其中羊种布鲁氏菌占多数(95.45%);20株布鲁氏菌分离株MLST基因型为ST8,1株为ST17型,1株为新的ST型:ST99的gap基因与已报道的等位基因型不同,被定义为一个新的等位基因型gap(32)。MLVA-16分型为羊种和猪种2个种群,21株羊种布鲁氏菌分为17种基因型,1株猪种布鲁氏菌分为1种基因型,其中15种基因型为单分离株,3种基因型为共享基因型(共7株,占31.82%)。聚类分析显示福建分离株与内蒙古、辽宁、山东和广东地区存在4种共享基因型,均为羊种布鲁氏菌,其他部分菌株与外省菌株遗传距离较近。结论福建省布鲁氏菌主要流行株为ST8型,且MLVA-16分型显示呈高度基因多样性。MLST/MLVA作为传统生物学鉴定补充技术方法,可用于布鲁氏菌遗传多样性分析和分子流行病学溯源调查,以提高布鲁氏菌病监测能力。Objective To explore the types of species and molecular characteristics of human brucellosis in Fujian Province in 2018-2019 in order to control and prevent brucellosis. Methods Based on cultures of Brucella isolates and extraction of their genomic DNA, conventional biological methods and molecular biological methods(BCSP31-PCR, AMOS-PCR, MLST, and MLVA-16) were used to molecularly identify and type Brucella. The products of PCR amplification were detected using the LabChip GX II Touch microfluidic capillary electrophoresis system with the 24 DNA Extended Range LabChip. MLST sequence types(STs) were determined via sequence alignment, and the MLVA-16 genotypes were grouped into clusters using the software Bionumerics 6.6. Results Molecular testing of 22 isolates of human Brucella in Fujian Province from 2018 to 2019 yielded results that were basically in line with those of conventional typing. Two species(B. melitensis and B. suis) and two biovars(B. melitensis biovar 3, n=21;and B. suis biovar 3, n=1) were identified. The B. melitensis species(21, 95.45%) was dominant. Among the 22 Brucella isolates, the most frequent ST was ST8(20 strains, 90.91%), followed by ST17(1 strain, FJ2019013) and a newly-identified ST(ST99, 1 strain, FJ2018003). The gap gene of ST99 differed from existing alleles and was designated as a new allele, gap(32). MLVA-16 revealed 17 genotypes in 21 B. melitensis isolates and 1 genotype in 1 B. suis isolate. These included 15 unique and 3 shared genotypes(7 isolates, 31.82%). The similarity of the two species(B. melitensis and B. suis) was less than 20%. A comparison to 180 isolates from other provinces indicated that 4 genotypes of B. melitensis in Fujian were also found in Inner Mongolia, Liaoning, Shandong, and Guangdong. Some isolates were genetically similar to isolates from other provinces. Conclusion The current study found that the Brucella strain mainly prevalent in Fujian is ST8, and MLVA-16 revealed a high level of genetic polymorphism. MLST/MLVA, as a supplementary technique for c

关 键 词：布鲁氏菌 多位点序列分型(MLST) 多位点可变数目串联重复序列分析(MLVA-16) 福建省 

分 类 号：R378.5[医药卫生—病原生物学]