现代检验医学杂志2026，Vol.41Issue(3)：50-54,60,6.DOI:10.3969/j.issn.1671-7414.2026.03.009
人空肠弯曲菌FlaA蛋白重组表达与免疫血清制备以及间接ELISA检测血清抗体的方法建立及初步应用
Recombinant Expression of the Human Campylobacter jejuni FlaA Protein,Preparation of Immuniserum,and Establishment and Preliminary Application of an Indirect ELISA Assay for the Detection of Serum Antibodies
童欣 1赵霞 2陆康妍 2夏洁 2黄金林3
作者信息
- 1. 扬州大学江苏省人兽共患病学重点实验室,江苏 扬州 225000||宜兴市红十字会血站,江苏宜兴 214200
- 2. 宜兴市红十字会血站,江苏宜兴 214200
- 3. 扬州大学江苏省人兽共患病学重点实验室,江苏 扬州 225000
- 折叠
摘要
Abstract
Objective To express and purify the Campylobacter jejuni(C.jejuni)FlaA protein and to develop an ELISA diagnos-tic kit for detecting C.jejuni antibodies in humans.Methods Bioinformatic analysis of the C.jejuni FlaA protein was performed,and the recombinant plasmid pET-28a-flaA was constructed.The recombinant protein was expressed by transforming BL21(DE3)competent cells.Female New Zealand white rabbits were immunized to prepare the corresponding polyclonal antiserum.The op-timal coating concentration was screened,experimental conditions were optimized,and an indirect ELISA assay was established.The assay was then applied to serum samples from 58 patients with diarrhea and 697 healthy blood donors to evaluate the sensi-tivity and specificity of the method.Results The pET-28a-flaA recombinant plasmid was successfully constructed.The recom-binant protein obtained via the prokaryotic expression system exhibited high expression levels.Western blot analysis confirmed that the recombinant protein reacted strongly with positive sera and could serve as a target antigen.An optimized indirect ELISA protocol was established,and the test results were generally consistent with clinical findings,with specificity and sensitivity of 75%and 99.86%,respectively.Conclusions The C.jejuni FlaA recombinant protein is successfully expressed and purified,and an indirect ELISA method using FlaA as the coating antigen is established.关键词
空肠弯曲菌/FlaA蛋白/质粒重组/蛋白表达纯化/间接酶联免疫吸附试验/多克隆抗体Key words
Campylobacter jejuni/FlaA protein/recombinant plasmid/protein expression and purification/indirect ELISA/polyclonal antibody分类
医药卫生引用本文复制引用
童欣,赵霞,陆康妍,夏洁,黄金林..人空肠弯曲菌FlaA蛋白重组表达与免疫血清制备以及间接ELISA检测血清抗体的方法建立及初步应用[J].现代检验医学杂志,2026,41(3):50-54,60,6.
