81例女性淋病患者孔蛋白基因型分布及与相应多克隆抗体的反应特性

Distribution of porins genotypes and reactivity to polyclonal antibodies in 81 female patients with gonorrhea

目的:采用PCR和探针技术分析临床淋病奈瑟菌野生株孔蛋白基因的基因型,并和相应多克隆抗体进行反应以检测基因突变对抗原性的影响。方法:采用PCR技术从淋病奈瑟菌野生株中扩增出特征性的孔蛋白基因片段,再用探针检测相应的高变区情况,以区分不同的准种(亚型),最后分别与原核表达的标准菌株孔蛋白多克隆抗体进行反应,观察相应的滴度。结果:孔蛋白基因型为A型的有35例,其余46例为B型。孔蛋白A型基因型变化较小,与抗体的反应性较一致;孔蛋白B型基因型变化稍大,但不影响与多克隆抗体的反应活性,A型与B型存在交叉反应,但活性较低,滴度<1∶40。结论:通过分析不同孔蛋白基因型淋病奈瑟菌与相应多克隆抗体的反应情况,有助于考察孔蛋白原核表达产物的生物学效应,对于人工合成出能针对绝大多数临床基因变异菌株的探针用于检测或治疗性抗体的制备有重要意义。

Objective:To analyze the genotype of the porin gene in Neisseria gonorrhoea wild strains. Bacterial agglutination has been carried out to verify whether there is an antigenicity change caused by the porin gene mutation.Methods:Porin gene was amplified by PCR from wild strains and probes were used to detect the mutation in VRs to differentiate the isoforms. Different strains reacted with the multi-clonal antibody to observe their titre.Results:There were 35 cases of type-A porin genotype and 46 cases of type B within the 71 cases. Greater mutations were found in type B than in type A with no effect on antigen-antibody reactivity. Cross reactivity existed between A and B with low titre(less than 1∶40).Conclusion:It must greatly help to inspect the biological effects of porins expressed by prokaryotic expression system in analyzing different porin genotype strains’ reactivity with antibody. It is also of significance to produce antibody with diagnostic or remedial potential aiming at most of strains of different porin genotypes