CAJ | 학술논문

为了鉴别布鲁菌疫苗免疫和自然感染，建立鉴别布鲁菌 S2疫苗株与其他菌株的斑点杂交法。针对布鲁菌基因序列保守区 S2疫苗株与其他菌株的差异设计探针，同时根据差异部分设计引物进行 PCR扩增；PCR 产物经过回收、纯化、变性后固定在 NC 膜上，与探针杂交、显色。结果显示，PCR 引物能够对 S2疫苗株扩增出约330 bp 的核酸片段，对其他布鲁菌扩增出约360 bp 的核酸片段；探针只能和 S2疫苗株的PCR 产物杂交，最低检测到10 pg DNA，能够在10 h 内鉴别出布鲁菌 S2疫苗株。
위료감별포로균역묘면역화자연감염，건립감별포로균 S2역묘주여기타균주적반점잡교법。침대포로균기인서렬보수구 S2역묘주여기타균주적차이설계탐침，동시근거차이부분설계인물진행 PCR확증；PCR 산물경과회수、순화、변성후고정재 NC 막상，여탐침잡교、현색。결과현시，PCR 인물능구대 S2역묘주확증출약330 bp 적핵산편단，대기타포로균확증출약360 bp 적핵산편단；탐침지능화 S2역묘주적PCR 산물잡교，최저검측도10 pg DNA，능구재10 h 내감별출포로균 S2역묘주。
To Explore the dot blot hybridization for discrimination of Brucella live vaccine strain S2 from field strains,according to the difference of geneme between B.suis vaccine strain S2 and other strains in NCBI,specific primers and probes were designed and labeled by DIG high prime.PCR products were recov-ered,purified,denatured and fixed to the NC membrane,then hybridized with the probe labeled by DIG high prime and detected.330 bp fragment could be amplified from strains of vaccine strain S2,and 360 bp from other Brucella strains.The probe was specific and no cross hybridization with other Brucella strains. 10 pg DNA of Brucella live vaccine strain S2 could be discriminated in 10 h by the dot blot hybridization.