随州市羊种3型布鲁氏菌首发病例的病原学检测

Pathogenic detection of the first cases of B.melitensis type Ⅲ in Suizhou city

目的分离和鉴定布鲁氏菌,为病例的诊断、治疗及疫情的控制提供科学依据。方法虎红平板凝集试验(RBPT)、试管凝集试验(SAT)、酶联免疫吸附试验(ELISA)检测病人血清中特异性抗体,用血清凝集试验、噬菌体试验、PCR试验法对菌株进行分型鉴定。结果虎红平板凝集试验出现絮状抗原抗体结合物100%凝集为阳性;试管凝集试验结果为阳性1:200(+++);发病期血清IgM抗体阳性、IgG阴性;M5布鲁氏菌疫苗株和病人菌株提取的基因组进行扩增和电泳,均在223和731bp附近出现阳性条带,而711、976和285 bp处无条带,说明该病人分离出的菌株为布鲁氏菌属羊种布鲁氏菌;经CO2、硫化氢、染料抑菌、菌株表面抗原凝集、噬菌体裂解试验,鉴定该菌株为羊种3型。结论随州市布病首发病例病原体为羊种3型布鲁氏菌,应加强布病监测,采用RBPT、SAT、ELISA、PCR等方法可快速检测病原体,为临床诊断、疫情控制提供依据。

Objectives To isolate and identify Brucella and provide scientific evidence for diagnosis, treatment and con- trol of Brucellosis. Methods Rose-Bengal plate agglutination test （RBPT）, standard tube agglutination test（SAT）, enzyme linked immunosorbent assay （ELISA） were used to test specific antibody in the sera. Serum agglutination test, Bacteriophage test and polymerase chain reaction（PCR） were used to identify the type of stains. Results Flocculent antigen-antibody combo 100% agglutination appeared in RBPT was positive; The result of SAT was positive （1：200, ＋＋ ＋）; The serum IgM antibody was positive and IgG was negative during the course of disease; Genomes were extracted from M5 Brucella vaccine strains and the patient strain. After amplification and electrophoresis, all appeared positive banding around 223 and 731 bp fragments and no banding at 711,976 and 285 bp. It indicated that the strain isolated from the patient was B.melitensi~, After stained with CO2, H2S, bacteriostasis, strain surface antigen agglutination and phage cracking test, the strain was identified as B.melitensis type ]1I. Conclusion The pathogen of first case was B.melitensis type HI in Suizhou. The monitoring work of brucellosis be strengthened. RBPT, SAT,ELISA and PCR allowed rapidly detection of Brucella and they can provide evidence for clinical di- agnosis and control the prevalence of brucellosis.