一株单核细胞增生李斯特菌分离株的分子分型鉴定及其生物学特性  

Genotyping Analysis and Biological Characteristics of a Clinical Listeria monocytogenes Isolate

作　　者：辛永萍 单颖 夏叶[4] XIN Yongping;SHAN Ying;XIA Ye(Clinical Research Center,The Second Af filiated Hospital,Zhejiang University School of Medicine,Hangzhou 310009,China;Zhejiang Provincial Key Lab of Preventive Veterinary Medicine,College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;Large Instrument Management Platform,College of Animal Sciences,Zhejiang University,Hangzhou 310058,China;Institute of Animal Husbandry&Veterinary Sciences,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China)

机构地区：[1]浙江大学医学院附属第二医院临床研究中心,杭州310009 [2]浙江大学动物科学学院,浙江省动物预防医学重点实验室,杭州310058 [3]浙江大学动物科学学院大型仪器管理平台,杭州310058 [4]上海市农业科学院畜牧兽医研究所,上海201106

出　　处：《畜牧兽医学报》2020年第5期1101-1109,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31802214,31702250);上海市农业科学院学科领域建设项目(专项农科国推2019匹配-09)。

摘　　要：从上海某羊养殖场获得了一株单核细胞增生李斯特菌分离株CMG47。为了确定该单核细胞增生李斯特菌分离株的分子分型,了解其生物学特性,本研究通过多重PCR对该菌株进行谱系和血清型分析,利用多位点序列分型(MLST)方法鉴定其分子分型。采用PCR方法对主要毒力基因进行检测,并通过体外观察和荧光定量PCR对菌株溶脂溶血特性进行分析。将菌株通过腹腔注射ICR小鼠和静脉注射斑马鱼,测定其毒力。研究结果表明该分离株属于谱系Ⅰ,1/2b血清型;序列分型为ST619;携带prfA、inlA、inlB、plcA、plcB、mpl、actA、hly等主要毒力因子;体外无明显溶脂活性,溶血活性较弱;小鼠和斑马鱼试验均显示,该分离株属于强毒株,与强毒参考株EGDe的毒力相当(P>0.05)。该分离株的谱系/血清型为引起李斯特菌病的主要型别,拥有整套主要毒力因子,为单增李斯特菌强毒株。本研究为李斯特菌病散发病例的流行和传播特征分析提供了分子生物学基础,对建立健全李斯特菌监测体系和风险评估意义重大。An isolate of Listeria monocytogenes CMG47 was obtained from a goat farm in Shanghai.To identify the genotyping and biological characteristics of this isolate,multiplex PCR was performed in lineage identification and serotyping,multi-locus sequence typing(MLST)was used for genotyping.Major virulence factors were examined by PCR,hemolysis and phospholipase activity were analyzed by in vitro observation and real-time fluorescent quantitative PCR.The virulence was determined by intraperitoneal injection in ICR mice and intravenous injection in zebrafish.Study results revealed that the isolate belonged to lineage I,1/2b for its serotyping,ST619 for its genotyping,and carried major virulence factors such as prfA,inlA,inlB,plcA,plcB,mpl,actA and hly.It showed weak hemolysis but did not exhibit apparent phospholipase activity.The isolate was of high pathogenicity comparable to the reference virulent strain EGDe in mice and zebrafish models(P>0.05).The isolate belongs to the dominant serotyping and genotyping for Listeria infection,which carries major virulence factors and is highly pathogenic.This work provides the molecular basis for further analysis of the prevalence and distribution characteristics of sporadic listeriosis and is very important for the establishment of Listeria monitoring system and risk assessment as well.

关 键 词：单核细胞增生李斯特菌 多位点序列分型 毒力 

分 类 号：S852.61[农业科学—基础兽医学] R378.994[农业科学—兽医学]