7株O139霍乱弧菌CT基因阴性菌的生物学性状研究  

Study on biological characteristics of seven ctx negative Vibrio cholerae O139 strains

作　　者：于纪桂[1] 刘延清[1] 祁国明[1] 高守一[1] 章丽娟[1] 

机构地区：[1]中国预防医学科学院流行病学微生物学研究所

出　　处：《中华微生物学和免疫学杂志》1999年第2期89-93,共5页Chinese Journal of Microbiology and Immunology

基　　金："863"项目基金

摘　　要：目的拟从毒力基因ｃｔｘ阴性的Ｏ１３９霍乱弧菌中筛选疫苗候选株。方法用常规鉴定技术、Ｓｏｕｔｈｅｒｎｂｌｏｔｉｎｇ技术和兔肠段结扎试验检测了７株霍乱肠毒素（ＣＴ）基因阴性的Ｏ１３９霍乱弧菌的一般特性、毒性和毒性基因；通过小肠免疫家兔，用ＰＢＬ－ＥＬＩＳＡ、ＥＬＩＳＡ和杀弧菌抗体试验评价抗原性；用家兔主动保护试验和乳鼠被动保护试验研究保护效果。结果７株菌都具有Ｏ１３９霍乱弧菌的一般特性，均不引起肠段积液，毒力基因ｃｔｘ、ｚｏｔ、ａｃｅ和ＲＳ１探针杂交结果均为阴性。Ｂ１３和９４００１免疫后０～２８天血清抗体ＩｇＧ效价一直上升，而９４００２免疫后１４天最高；９４００２的血清杀弧菌抗体峰值最高。Ｂ１３免疫家兔后可保护１０５ＣＦＵ毒株的攻击，９４００１和９４００２免疫后可保护１０５～７ＣＦＵ毒株的攻击，９４００２加强免疫后能保护１０７ＣＦＵ毒株的攻击。Ｂ１３免疫血清１４稀释后对乳鼠的保护率为２０％，９４００１和９４００２的免疫血清１１６稀释后的保护率为５０％。结论７株Ｏ１３９霍乱弧菌属于毒力基因遗传单元缺失，无毒或毒性低，其中９４００２的免疫原性和保护效果较好。Objective Vibrio cholerae O139, which was found in many countries and regions in the world in recent years, has been identified as the source of a cholera like disease. So it is necessary to develop an effective vaccine for prevention of the epidemic. Seven Vibrio cholerae O139 strains whose ctx is negative were studied. Methods The basic characteristics were detected with routine microbiology test and biochemical test, Southern blotting, and ligated rabbit ileal loop test. The immunogenicity was evaluated with vibriocidal antibodies test, ELISA and PBL ELISA, and the protective efficacy with active protective test in rabbit model and passive protective test in infant mouse model. Results All of the 7 strains had the general characteristics of Vibrio cholerae O139, were negative with ctx, zot, ace and RS1, and could not evoke ileal loop response after being administered to rabbits, The serum titers of IgG and vibriocidal antibody of rabbits, immunized with B13 and 94001 respectively, went up from day 0 to day 28, but with 94002, a peak appeared on the 14 th day, and its vibriocidal antibody titer was the highest among the 3 strains. The rabbits immunized with B13 were protected against the challenge of 10 5 CFU virulent strains, those immunized with 10 5 7 CFU of 94001 or 94002, respectively, and those immunized and boosted with 94002,10 7 CFU. In infant mouse model, to the strain B13, the immune serum diluted by 4 fold had 20% protective rate, to 94001 and 94002, diluted by 16 fold had 50% protective rate. Conclusions The results prove that all of the 7 strains lack the whole virulence cassette and have little virulence. The strain 94002 exhibites not only the best immunogenicity and protective efficacy, but also immune memory.

关 键 词：霍乱弧菌 CT基因阴性菌株 免疫性 

分 类 号：R378.3[医药卫生—病原生物学]