摘要
为了开发布鲁氏菌病新型标记疫苗,本研究对流产布鲁氏菌基因缺失株RB6的培养特性、染色特性、凝集特性、稳定性及小鼠体内毒力和免疫保护力进行了系统鉴定,旨在阐明该菌株具备的生物学特性。通过对亲本菌株和RB6的比较,发现固体培养基上RB6菌株单菌落可被结晶紫染成紫色,RB6菌株液体培养物可与0.1%吖啶黄染料以及抗布鲁氏菌粗糙型抗体发生凝集反应,证明该菌株为粗糙型。将RB6菌株在体外连续传代培养20次和牛体内连续5次继代,检测结果证明其表型未发生变化,说明该菌株遗传稳定性良好。通过小鼠体内试验发现该菌株毒力显著降低,并对流产布鲁氏菌强毒菌株2308攻毒的免疫保护力与现有疫苗A19接近。本研究结果表明,流产布鲁氏菌基因缺失株RB6为粗糙型菌株,毒力较低、安全性高、遗传性状稳定,并具有良好的免疫保护效力,有望开发成为动物布鲁氏菌病粗糙型标记疫苗。
To develop a novel marker brucellosis vaccine,the gene-deleted mutant RB6 derived from Brucella abortus was used in this paper to examine its essential properties.By comparing growth characteristics,staining properties,agglutination identities,stability,virulence and immunological protection between RB6 and its parent strain,it was found that the growth characteristics of RB6 was similar to the parent strain,but the colonies on TSA became to the violet in the crystal violet staining test,and the cultural suspension was agglutinated with sera against Brucella rough-type strain,which showed that RB6 was a rough phenotype.It was also demonstrated that the phenotype and virulence of RB6 was stable when it was subcultured for 20 passages in vitro and then for 5 passages in heifers.In the mice model,RB6 was attenuated and induced the similar immunological protection levels as A19 vaccine strain.In a word,RB6 was a rough-type mutant with low virulence,high genetic stability and good protection efficacy,which could be employed to prepare new marker vaccines using for the prevention and control of animal brucellosis.
出处
《中国畜牧兽医》
CAS
北大核心
2012年第4期174-177,共4页
China Animal Husbandry & Veterinary Medicine
基金
国家973计划(2010CB530202)
关键词
流产布鲁氏菌
粗糙型疫苗
稳定性
毒力
免疫效力
Brucella abortus
rough-type vaccine
stability
virulence
immune efficacy
