霍乱弧菌小川型在人胆汁中存活和变异及其血清型别转换的分子生物学研究  

Molecular biological study on the survival,variation and serotypeconversion of Vibrio Ogawa strains in human bile (in Vitro)

作　　者：盛清[1] 凌志强[2] 王满琴[2] 郑伟[2] 何浙生[2] 

机构地区：[1]浙江理工大学,杭州310018 [2]浙江省医学科学院,杭州310013

出　　处：《中国卫生检验杂志》2004年第6期667-670,共4页Chinese Journal of Health Laboratory Technology

摘　　要：〔目的〕追踪观察霍乱弧菌小川型流行株在离体人胆汁样本中的存活和L型变异状况 ;探索小川型在其抗体作用下对转换为稻叶型的发生频度和其对Ctx基因的影响 ;为改善当前霍乱防治工作提供实验依据。〔方法〕对保存染菌胆汁样本进行细菌和L型测数、鉴定。以抗体软琼脂法诱导血清型别转换 ;以蔗糖半固体平板检测细菌趋化性。用PCR和测序法比较转型前后的肠毒素基因改变。〔结果〕在 2份染菌人胆汁中 ,发现 1份存活达 2 2年 ,其总菌数为 2 .5× 10 3 cfu/ml,其中L型构成比为 32 %。 3株小川型流行株在人胆汁及抗体作用下 ,转换为稻叶型发生频度为 2 %～ 6 % ;酶切电泳显示转型前后其Ctx基因图型相似 ,经CtxAB测序后 ,差异≤ 7Nt(7/12 2 6 ) ,显示有一定的变异。 3株转型前后细菌的化学趋化性未见显著改变。〔结论〕为霍乱弧菌在某些患者胆囊中长期存活而成为慢性带菌者提供了实验依据。Objective]To pursue and observe the surviving and L-form variation of the epidemic strains of Vibrio cholerae Ogawa in human′s bile in vitro; to detect the frquency of the serotype Ogawa strain′s conversion into serotype Inaba under its antisera′s action and the influence of serotype conversion on gene Ctx; To supply experimental data for improving the current prevention and treatment of cholera. [Methods] Identify and take count of the germs and L-form of the samples,the bile contaminated with bacteria; Contaminate the bile with V.Cholerae E1 Tor biotype Ogawa strains and take count of them;Induce the serotype conversion by soft agar containing antisera; Examine the bacterial chemotaxis with fore-and-aft serotype conversion of V.Cholerae E1 Tor biotype Ogawa strains with sucrose semisolid petri dish; To compare the entrotoxin gene after the serotype conversion with PCR and determining its sequence.[Results] We discover one of two bile samples contaminated by V. Cholerae has been surviving for 22 years, and its total bacterial count is 2.5ⅹ10 3/ml, in which the constituent ratio of L-form germs is 32%.The frequency of conversion into serotype Inaba of three prevailing strains of V.cholerae Ogawa is 2%～6%.The electrophoresis after enzyme′s detaching reveals that the patterns of gene Ctx are similar, and the difference is ≤10Nt (10/1226) after the sequence determination of CtxAB.It shows some existed variations,but the bacterial chemotoxis of three strains with fore-and-aft serotype conversion has not changed significantly.[Conclusions] The results also provided the new experimental data for improving the current prevention and treatment of cholera.

关 键 词：胆汁 霍乱弧菌 L型 存活 抗体 型别 X基因 变异 血清型 流行株 

分 类 号：R378[医药卫生—病原生物学] R735[医药卫生—基础医学]