甘肃省靖远县绵羊血液内分离的布鲁菌菌株种型鉴定

Identification of Brucella strain isolated from sheep blood in Jingyuan County Gansu Province

目的对分离自甘肃省靖远县绵羊血液的布鲁菌菌株进行种型鉴定，为布鲁菌病（布病）疫情的防治提供科学依据。方法2016年，在靖远县布病患者家中．采集疑似感染布病的绵羊血液，接种至双相血培养瓶中进行增菌培养，再分别采用传统的生物分型法、布鲁菌属特异性表面蛋白基因31-PCR（BSCP31-PCR）和牛、羊、绵羊附睾、猪种布鲁菌特异性PCR（AMOS—PCR）3种方法对增菌培养的待测菌株进行鉴定。结果分离菌株经传统的生物分型法鉴定，在硫堇、复红染料中均生长，在单相特异性血清A和M中都为阳性，Bk2噬菌体对待测菌株裂解，Tb、Wb噬菌体则不裂解；BSCP31-PCR扩增结果为223bp条带，与布鲁菌属一致：AMOS—PCR扩增结果为731bp条带，与羊种布鲁菌菌株一致。结论采用传统生物分型法，结合分子生物学方法鉴定，在甘肃省靖远县绵羊血液内分离的布鲁菌菌株为羊种3型布鲁菌。

Objective To identify the Brucella strains isolated from sheep blood in Jingyuan County Gansu Province, and to provide scientific basis for Brucellosis control and prevention. Methods In 2016, blood samples from suspected cases of sheep were collected in the home of Brucellosis patient of Jingyuan County, and cultured with vaccination double blood bottles via the enrichment method, and the Brucella strains isolated from sheep were identified by traditional bio-typing, genus specific Brucella surface protein 31 PCR （BSCP31-PCR） and Brucella abortus, Brucella melitensis, Brucella ovis, Brucella suis species-specific PCR （AMOS-PCR）, respectively. Results The isolated strain was identified using the traditional biological classification method, bacteria could grow in thionine and fuchsin dyes, A and M factors serum agglutination tests were positive, Bk2 phage lysis test was positive, but Tb and Wb phage lysis tests were negative; and the results of BSCP31-PCR of tested strain was 223 bp band same to Brucella; the results of AMOS-PCR was 731 bp band same to Brucella melitensis. Conclusion Conventional bio-typing test assisted with molecular techniques has confirmed that the isolated Brucella strain from sheep blood is Brucella melitensis type 3 in Jiugyuan County Gansu Province.