绵羊李斯特菌病病原诊断及其生物学特性研究  

Diagnosis of Listeria monocytogenes NTSN

作　　者：王国梁[1] 殷月兰[1] 焦库华[1] 张小荣[1] 付红[1] 高云飞[1] 贾艳艳[1] 焦新安[1] 

机构地区：[1]扬州大学,江苏省人兽共患病学重点实验室,扬州225009

出　　处：《中国人兽共患病学报》2013年第7期639-645,共7页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.31101841);江苏省科技支撑计划(BE2012367);江苏省自然科学基金(No.BK2011446)资助项目资助~~

摘　　要：目的单核细胞增生性李斯特菌(Lm)是引起人兽共患李斯特菌病的病原菌,该菌感染宿主中枢神经系统是导致高死亡率的主要原因。本研究对农场暴发的呈现神经症状的绵羊李斯特菌病病例进行病原诊断,进而对其病原学特性开展相关研究。方法利用选择培养基和鉴别培养基进行细菌的分离和鉴定,借助多重PCR方法和Lm诊断血清对分离株的血清型进行研究,并测定了该菌株的溶血活性和药物敏感性,基于毒力基因actA的遗传谱系进行分型,最后对其LD50进行了测定。结果从绵羊脑实质组织中成功地分离到一株血清型为4b的Lm菌株NTSN,其溶血效价为24,LD50为104.30 CFU,是具脑侵袭性的强毒菌株,可用青霉素类药物治疗。遗传谱系分析表明Lm NTSN可能属于高侵袭性的流行克隆。结论 LmNTSN的分离鉴定及其生物学特性的研究,为深入探讨Lm的致病机理和预防控制奠定了基础。Listeria monocytogenes （Lm） is an important pathogen that can cause zoonosis, its invasion and infection central nervous system account for the high mortality associated with listeriosis. In this study, an outbreak of sheep listeriosis with neurological symptoms was diagnosed and then characterization of its pathogenicity was investigated. Firstly, a listerial strain named Lm NTSN was isolated by selective medium. Additionally, multi-PCR and the serum agglutination test against Listeria monocytogenes were conducted to determine its serotype. The hemolysis activity was determined with sheep erythrocytes, the drug susceptibility of Lm NTSN was carried out by disc diffusion test. Its virulence was evaluated on BALB/c mouse model. Phylogenetic analysis based on the partial actA sequences was performed. Results indicated that a Listeria monocytogenes 4b strain NTSN from sheep cerebral parenchyma was isolated and identified. Its hemolysis titers reached 24, LD50 to BALB/c mouse was 104.30 CFU, so Lm NTSN was a highly pathogenic, brain invasive strain. Penicillin can be used to treat this case of listeriosis. The genetic analysis also indicated that the serotype 4b Lm may belong to a highly invasive epidemic clone. In conclusion, the biological characterization of Lm NTSN laid the foundation on research work about pathogenic mechanism, as well as prevention and control of sheep listeriosis.

关 键 词：单核细胞增生性李斯特菌 羊李斯特菌病 分离鉴定 毒力 

分 类 号：S855.1[农业科学—临床兽医学]