CAJ | 학술논문

通过构建标记疫苗株来解决流产布氏杆菌(B.abortus)鉴别诊断方面的缺陷,本研究以bp26基因作为重组靶住点,S19为亲本,利用bp26基因ORF外侧序列作为同源重组序列,卡那霉素抗性基因(Kanr)为抗性筛选标记,通过双交叉重组筛选获得bp26基因缺失突变的重组S19株,命名为S19-△26.小鼠感染结果表明,突变株S19-△26的残留毒力与亲本株S19相比较没有发生明显改变,康复时间约为15周,突变株S19-△26、亲本株S19和B.abortus强毒株S544接种小鼠后的第3周能检测出"O"抗原的特异性抗体,而第6周开始S19和S544接种小鼠BP26特异性抗体明显升高,S19-△26接种的小鼠一直没检测到BP26特异性抗体.小鼠免疫保护试验显示,脾脏分离CFU数比空白对照要低310g10,S544攻击后脾脏细菌分离数表明突变株具有与亲本疫苗株免疫保护性无明显差异.结果表明,S19-△26免疫能够通过血清学方法与野生型B.abortus感染后的免疫反应相区别,具备作为标记疫苗的潜力.
통과구건표기역묘주래해결유산포씨간균(B.abortus)감별진단방면적결함,본연구이bp26기인작위중조파주점,S19위친본,이용bp26기인ORF외측서렬작위동원중조서렬,잡나매소항성기인(Kanr)위항성사선표기,통과쌍교차중조사선획득bp26기인결실돌변적중조S19주,명명위S19-△26.소서감염결과표명,돌변주S19-△26적잔류독력여친본주S19상비교몰유발생명현개변,강복시간약위15주,돌변주S19-△26、친본주S19화B.abortus강독주S544접충소서후적제3주능검측출"O"항원적특이성항체,이제6주개시S19화S544접충소서BP26특이성항체명현승고,S19-△26접충적소서일직몰검측도BP26특이성항체.소서면역보호시험현시,비장분리CFU수비공백대조요저310g10,S544공격후비장세균분리수표명돌변주구유여친본역묘주면역보호성무명현차이.결과표명,S19-△26면역능구통과혈청학방법여야생형B.abortus감염후적면역반응상구별,구비작위표기역묘적잠력.
Brucella abortus strain S19 played an important role in preventing and monitoring brucellosis with high protective rate. S19-△26, a Mutant bp26 gene of B. Abortus S19 strain was constructed by replacing the bp26 gene of attenuated S19 with kanamycin resistence gene. Compared with Si9, mutant S19-△26 possessed the same recover time (15 weeks) and residual virulence in mice, However it did not induce anti-bp26 antibody production in Mice, which would differentiate animals infected with S19-△26 from those infected by S19 and S544. The mutant S19-△26 possesses the same protective ability as S19, which could reduce CFU number of strain S544 infection in mice by 31og10. Altogether, these results indicate that B. Abortus S19-△26 could be a promising vaccine strain in preventing brucellosis.