CAJ | 학술논문

目的对青海省2013年分离的布鲁杆菌菌株进行种型鉴定.方法 2013年7-8月,在青海省13个布鲁杆菌病(简称布病)病区收集60份人、畜布病样本,包括33份羊(牛)流产羔、27份布病抗体阳性患者血样.取羊(牛)流产羔的肝脏、脾脏、肺组织和布病患者血标本进行病原分离培养,经革兰染色、硫堇和复红染料抑菌试验、A和M因子血清凝集试验、噬菌体裂解试验鉴定后,得到待测菌株;采用AMOS-PCR、多重PCR和多位点可变数量串联重复序列分析(MLVA)-16分型方法对待测菌株进行种型鉴定.结果 60份人、畜布病样本经病原分离培养鉴定得到3株待测菌株.菌落形态观察呈单个散状排列,湿润、圆形、稍隆起,表面光滑、无色透明;革兰染色均为阴性;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;Bk噬菌体对3株待测菌株裂解,Tb、Wb噬菌体则不裂解;AMOS-PCR与多重PCR的扩增结果与羊种布鲁杆菌M5一致;3株待测菌株MLVA-16分型的基因型均与羊种菌株聚类在一起.结论采用布鲁杆菌分型方法,结合分子生物学方法鉴定,分离的布鲁杆菌菌株为羊种3型布鲁杆菌.
목적 대청해성2013년분리적포로간균균주진행충형감정.방법 2013년7-8월,재청해성13개포로간균병(간칭포병)병구수집60빈인、축포병양본,포괄33빈양(우)유산고、27빈포병항체양성환자혈양.취양(우)유산고적간장、비장、폐조직화포병환자혈표본진행병원분리배양,경혁란염색、류근화복홍염료억균시험、A화M인자혈청응집시험、서균체렬해시험감정후,득도대측균주;채용AMOS-PCR、다중PCR화다위점가변수량천련중복서렬분석(MLVA)-16분형방법대대측균주진행충형감정.결과 60빈인、축포병양본경병원분리배양감정득도3주대측균주.균락형태관찰정단개산상배렬,습윤、원형、초륭기,표면광활、무색투명;혁란염색균위음성;재류근、복홍염료중균생장;재단상특이성혈청A화M중도위양성;Bk서균체대3주대측균주렬해,Tb、Wb서균체칙불렬해;AMOS-PCR여다중PCR적확증결과여양충포로간균M5일치;3주대측균주MLVA-16분형적기인형균여양충균주취류재일기.결론 채용포로간균분형방법,결합분자생물학방법감정,분리적포로간균균주위양충3형포로간균.
Objective To identify three suspected strains of Bruce lla isolated in Qinghai Province in 2013.Methods From July to August in 2013,60 brucellosis samples of people and animals from 13 brucellosis endemic areas were collected in Qinghai Province,including 33 cattle and sheep abortion lambs and 27 brucellosis antibody positive blood samples of patients.The isolation and culture of pathogen test were carried out in the tissues of liver,spleen and lung from cattle and sheep abortion lambs and the serum samples from patients with brucellosis.After biochemical detection with gram staining,thionine and fuchsin dyes bacteriostasis test,A and M factors serum agglutination test and phage lysis,the tested strains were got.AMOS-PCR,multiplex-PCR and multiple-locus variable-number tandem-repeat analysis(MLVA)-16 were conducted to identify the types of the measured strains.Results Three tested strains were obtained from 60 brucellosis samples of people and animals by isolation and culture of pathogen test.The colonies were arranged in bulk,moist,round,slightly elevated,smooth,and colorless; the gram staining was negative; bacteria could grow in thionine and fuchsin dyes; A and M factors serum agglutination test were positive; Bk phage lysis test was positive,but Tb and Wb phage lysis tests were negative; the results of AMOS-PCR and multiplex-PCR of 3 tested strains were same to Brucella melitensis M5 ; the genetypes were clustered together with Brucella melitensis by MLVA-16 method.Conclusion Conventional typing test assisted with molecular techniques has confirmed that the isolated Brucella strains are Brucella melitensis type 3.