肺炎嗜衣原体三种不同抗原的临床诊断价值评价  

Comparison of three different novel Chlamydophila pneumonia recombinant antigens for serodiagnosis

作　　者：陈虹亮[1] 胡旃[1] 周洲[1] 曾焱华[1] 代国知 林应标 吴移谋[1] 

机构地区：[1]南华大学病原生物研究所,衡阳421001 [2]郴州第一人民医院检验科

出　　处：《中华检验医学杂志》2010年第2期138-142,共5页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目（30870134/C010902）

摘　　要：目的研究肺炎嗜衣原体3种重组抗原Cpn0146、Cpn0147及Cpn0308应用于肺炎嗜衣原体感染血清学诊断中的价值。方法构建pGEX6p-2／Cpn0146、Cpn0147、Cpn0308重组质粒，并诱导表达重组蛋白，采用间接ELISA法和免疫印迹法（Western—blot）分析其免疫原性及其免疫反应性。建立分别以3种重组蛋白为包被抗原的间接ELISA法，与肺炎嗜衣原体IgG商品诊断试剂盒平行检测183份呼吸道感染患者的血清标本及32份沙眼衣原阳性患者血清标本，比较各方法检出准确性及阳性率。结果GST-Cpn0146、Cpn0147、Cpn0308免疫兔血清中特异性IgG抗体效价滴度分别达1：6400、1：12800和1：12800以上；以3种重组蛋白为包被抗原的ELISA法准确性分别为92．3％、94．5％、96．7％。各方法检测出阳性率结果显示，Cpn0146组的阳性率为38．8％（71／183）、与肺炎嗜衣原体IgG试剂盒的阳性率比较差异有统计学意义（χ^2=12．07，P〈0．05）；Cpn0147阳性率40．9％（75／183），与肺炎嗜衣原体IgG试剂盒的阳性率比较差异有统计学意义（χ^2=8．10，P〈0．05）；Cpn0308阳性率44．8％（75／183），与肺炎嗜衣原体IgG试剂盒的阳性率比较差异无统计学意义（χ^2=0．57，P〉0．05）。结论Cpn0308在肺炎嗜衣原体感染血清学诊断实验中表现出良好的准确性（96．2％）和较高的阳性率（44．8％），可望用于进一步研制肺炎嗜衣原体诊断试剂盒。Objective To evaluate three different Chlamydophila pneumoniae recombinant antigens for use in Chlamydophila pneumoniae serodiagnosis. Methods The recombinant plasmids pGEX6p-2/ Cpn0146, Cpn0147 and Cpn0308 were constructed and expressed as GST fusion proteins. The immunogenicity and the immunocompetence of these recombinant protein were analyzed by Western-blot and indirect ELISA. A total of 183 sera samples of patients with respiratory tract infection and 32 sera samples of patients with Chlamydia trachomatis infection were detected with indirect ELISA coated microwell plates with the purified recombinant proteins comparing with SeroCP-TM IgG ELISA kits. The positive recognition rate, sensitivity and specificity of each method were analyzed. Results GST-Cpn0146, Cpn0147 and Cpn0308 were obtained after expression and purification. The titers of the specific IgG antibodies against Cpn0146, Cpn0147 and Cpn0308 were higher than 1：6 400, 1：128 00 and 1：128 00, respectively. When the indirect ELISA was developed to detect the IgG antibody against Chlamydophila pneumoniae in 183 samples, the concordance rate between the indirect ELISA test and SeroCP-TM IgG ELISA kits were 92. 3% （CpnO146）, 94. 5% （ Cpn0147 ） and 96.7% （ Cpn0308 ）, respectively. The recombinant Cpn0146, Cpn0147 and Cpn0308 were recognized by 71 （38.8% positive recognition rate）, 75 （40. 9% ）, and 82 （44. 8% ） samples, respectively. The recombinant antigen-based detection assays displayed 〉 97% of detection specificity and 〉 87% of sensitivity. Conclusion GST-Cpn0308 shows a better sensitivity and specificity, which suggests it could be used for developing serodiagnosis kits of Chlamydophila pneumoniae infection.

关 键 词：衣原体 肺炎 抗原 细菌 重组蛋白质类 血清学试验 

分 类 号：R563.1[医药卫生—呼吸系统]