甘肃省肃北蒙古族自治县喜马拉雅旱獭疫源地鼠疫耶尔森菌分子流行病学特征研究  

Molecular epidemiological characteristics of Yersinia pestis in Marmota himalayana plague foci in Subei Mongolian Autonomous County,Gansu Province

作　　者：郭丽民[1] 张晓玲 黄燕妍 赵存寿 张成鑫 付国明 GUO Li-min;ZHANG Xiao-ling;HUANG Yan-yan;ZHAO Cun-shou;ZHANG Cheng-xin;FU Guo-ming(Department of Plague Control and Prevention,Gansu Provincial Center for Disease Control and Prevention,Lanzhou 730020,China;Subei Mongolian Autonomous County Maternal and Child care Hospital,Subei 736300,China;Subei Mongolian Autonomous County center for Disease Control and Prevention,Subei 736300,China)

机构地区：[1]甘肃省疾病预防控制中心鼠疫防制科,兰州730020 [2]肃北蒙古族自治县妇幼保健院,肃北736300 [3]肃北蒙古族自治县疾病预防控制中心,肃北736300

出　　处：《中国人兽共患病学报》2025年第2期158-163,170,共7页Chinese Journal of Zoonoses

基　　金：甘肃省自然科学基金(No.22JR11RA185)。

摘　　要：目的 研究肃北县鼠疫菌差异区段(DFR)、规律成簇的间隔短回文重复序列(CRISPR)及可变数目串联重复序列(VNTR)的基因多态性特征,指导突发鼠疫疫情的溯源。方法 对1973-2017年间肃北县鼠疫疫源地不同动物宿主分离的89株鼠疫菌进行培养,提取DNA。分别利用DFR、CRISPR和MLVA基因分型的引物进行PCR反应,通过琼脂糖电泳观察扩增产物有无,与中国鼠疫菌DFR数据库比较确定鼠疫菌DFR基因型;通过PCR产物测序,与在线鼠疫菌CRISPR数据库比对确定CRISPR基因型;通过毛细管电泳计算每株鼠疫菌VNTR的重复数,基于肃北县89株与11株青海省喜马拉雅旱獭疫源地的鼠疫菌VNTR重复数应用BioNumerics7.6构建最小生成树。结果 肃北县89株鼠疫菌可分为6个DFR基因型,分别为8型、7型、1b型、5型、32型和44型,其中8型为肃北县鼠疫疫源地主要基因组型。肃北县89株鼠疫菌经CRISPR分型,可分为3个基因簇和3个基因型,其中Ca35′为肃北县鼠疫疫源地的主要基因簇,基因型为26′,主要分布在肃北蒙古族自治县党城湾镇、鱼儿红乡和石包城乡;Ca7和CaΔ5′为肃北县鼠疫疫源地的次要基因簇,基因型为22和24型,分布在党城湾镇、鱼儿红乡和石包城乡。MLVA聚类分析,肃北县鼠疫菌主要分为3个簇,分别为党城湾镇马场簇、鱼儿红乡簇和党城湾镇簇。结论 DFR、CRISPR和MLVA基因分型方法均能将肃北县鼠疫菌分为主要基因型和次要基因型,具有显著的地区分布特征,基因型多样性较高,种群特征复杂,尤其在鼠疫疫情溯源时给予重视。This study was aimed at determining the genetic characteristics of Yersinia pestis in Subei County through differential region(DFR),clustered regularly interspaced short palindromic repeats(CRISPR),and variable number tandem repeat(VNTR)analyses,to guide the tracing of plague outbreaks.The DNA of 89 Yersinia pestis strains isolated from various animal in foci of Subei County from 1973 to 2017 was extracted.Primers for genotyping by DFR,CRISPR,and MLVA were used in PCR,and agarose electrophoresis was used to determine whether the amplified products were present.Genotypes were determined through comparison against the DFR database of Yersinia pestis in China.The PCR products were sequenced and compared against the online CRISPR database for Yersinia pestis to determine the genotype.The number of VNTR repeats in each strain was calculated through capillary electrophoresis,and the minimum spanning tree was constructed with BioNumerics 7.6 according to the numbers of VNTR repeats from 89 Yersinia pestis strains in Subei County and 11 strains from a Marmota himalayana focus in Qinghai Province.The Yersinia pestis strains in Subei County were divided into six main genotypes by DFR:8,7,1b,5,32,and 44.The Yersinia pestis strains were divided into three gene clusters and three genotypes by CRISPR.Ca35′was the main gene cluster in Subei County;the genotype was 26′;and the distribution was primarily in Dangchengwan Town,Yuerhong Township,and Shibaocheng Township.Ca7 and CaΔ5′comprised secondary gene clusters,with genotypes 22 and 24,and were distributed in Dangchengwan Town,Yuerhong Township,and Shibaocheng Township.The Yersinia pestis strains in Subei County were divided primarily into three clusters:the Dangchengwan Machang cluster,Yuerhong Township cluster and Dangchengwan Town cluster.Therefore,the Yersinia pestis strains in Subei County,divided into major and minor genotypes according to DFR,CRISPR and MLVA,showed different regional distribution characteristics,highly diverse genotypes,and complex population char

关 键 词：肃北县 差异区段 规律成簇的间隔短回文重复序列 多位点可变数目串联重复序列 

分 类 号：R378.6[医药卫生—病原生物学]