肺炎支原体P116基因FP2片段的免疫优势和细胞黏附活性  

Immunodominance of FP2 fragment of P116 gene of Mycoplasma pneumoniae and cell adhesion activity

作　　者：林康 文志 周畅[1] 李文丽 吴焕武 刘丹艳 朱玉林[3] 王林定[1] Lin Kang;Wen Zhi;Zhou Chang(Dept of Microbiology,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学微生物学教研室,合肥230032 [2]阜阳师范大学,阜阳236000 [3]安徽医科大学第一附属医院儿科,合肥230032

出　　处：《安徽医科大学学报》2021年第8期1268-1272,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81271837);安徽高校自然科学研究项目(编号:KJ2012A161);安徽省自然科学基金项目(编号:1708085MH193);安徽医科大学基础与临床合作研究提升计划(编号:2019xkjT024);2019年度安徽医科大学校科研基金项目(编号:2019xkj020)。

摘　　要：目的通过对肺炎支原体P116基因FP2段的453 bp片段进行了克隆表达,探讨肺炎支原体P116蛋白对A549细胞的黏附性。方法肺炎支原体感染者血清和肺炎支原体抗体用Western blot和ELISA检测纯化的P116蛋白片段的免疫原性及效价。P116-FP2多克隆抗体和A549细胞黏附试验用免疫荧光评价肺炎支原体的黏附、黏附抑制。结果 P116-FP2蛋白具有免疫原性,用该蛋白构建的ELISA方法具有较高的敏感性和特异性。制备的P116-FP2抗体可阻断肺炎支原体黏附A549细胞。随着多克隆抗体效价的增加,肺炎支原体对A549细胞的黏附率明显降低。结论初步证明P116-FP2蛋白具有免疫反应性。P116-FP2多克隆抗体能够抑制肺炎支原体对A549细胞的黏附。Objective To clone and express the 453 bp fragment of P116 ge-ne FP2 of Mycoplasma pneumoniae,and to investigate the adhesion of P116 protein to A549 cells.Methods The immunogenicity and titer of purified P-116 protein fragment were detected by Western blot and ELISA in serum and antibody of Mycoplasma pneumoniae infected.Immunofluorescence was used to evaluate the adhesion and adhesion inhibition of Mycoplasma pneumoniae to A549 cells by P116-FP2 polyclonal antibody.Mycoplasma pneumoniae adhesion,adhesion inhibition and surface exposure were evaluated using the anti-P116 polycl-onal antibody and A549 cells adhesion assay.Results The results showed that P116-FP2 had immunoreactivity,and the ELISA method established with this protein had high sensitivity and specificity.The P116-FP2 antibody was prepared to block the adhesion of Mycoplasma pneumoniae to A549 cells.With the increase of titer of polyclonal antibody,the adhesion rate of Mycoplasma pneumoniae to A549 cells decreased significantly.Conclusion The P116-FP2 protein is proven to be immunoreactive preliminarily.The P116-FP2 polyclonal antibody can inhibit the adhesion of Mycoplasma pneumoniae to A549 cells.

关 键 词：肺炎支原体 P116蛋白 A549细胞 多克隆抗体 

分 类 号：R375.2[医药卫生—病原生物学]