CAJ | 학술논문

目的建立一步法PCR快速鉴定布鲁氏菌种及种内部分生物型.方法设计6对引物,建立一步法PCR反应,将布鲁氏菌6个种和种内某些生物型进行鉴定,应用该方法检测了27株布鲁氏菌参考菌株和239株临床分离的布鲁氏菌,并与生物学分型方法进行比较.结果通过一步法PCR能够将待检菌株准确的鉴定到布鲁氏菌种,对于猪种可以定位到生物型和疫苗株；对于牛种可以鉴定到牛种生物型1、3、4 与2、5、6、7、9两组生物型；对于羊种可以鉴定到1与2、3两组生物型和羊种疫苗株.生物学分型方法和PCR方法准确鉴定到种水平的符合率分别为98.33％和100％.结论一步法PCR是一种快速、特异、简便而低费用的鉴定布鲁氏菌种的分子分型方法.
목적 건립일보법PCR쾌속감정포로씨균충급충내부분생물형.방법 설계6대인물,건립일보법PCR반응,장포로씨균6개충화충내모사생물형진행감정,응용해방법검측료27주포로씨균삼고균주화239주림상분리적포로씨균,병여생물학분형방법진행비교.결과 통과일보법PCR능구장대검균주준학적감정도포로씨균충,대우저충가이정위도생물형화역묘주；대우우충가이감정도우충생물형1、3、4 여2、5、6、7、9량조생물형；대우양충가이감정도1여2、3량조생물형화양충역묘주.생물학분형방법화PCR방법준학감정도충수평적부합솔분별위98.33％화100％.결론 일보법PCR시일충쾌속、특이、간편이저비용적감정포로씨균충적분자분형방법.
Objective To develop a one-step PCR assay for rapid discrimination of six Brucella species and some intraspecific biovars.Methods Using 6 pairs of primers in one-step PCR to differentiate six classical Brucella species and some biovar in ordinary PCR instrument.The tested strains including 27 reference strains of six Brucella species and 239 Brucella strains were estimated by the PCR assay and biological identification methods.Results The six Brucella species could be precisely differentiated by the onestep PCR assay from the tested strains.Five biovars and vaccine strain of B.suis species could be determined,and biovars 1,3,4 and biovars 2,5,6,7,9 of B.abortus species could be identified at the level of their biovar,moreover,biovars 1,2 and 3,and vaccine strain Rev 1 of B.melitensis species were also discriminated at the biovar and strain level.The accurate rates of the biological identification method and the PCR assay were 98.33％ and 100％ respectively.Conclusion One-step PCR assay was a rapid,specific,and low cost method for identification of Brucella species and discriminating biovars in ordinary PCR instrument.