肺炎衣原体主要外膜蛋白重组抗原制备与应用的研究

Preparation and application of chalamydia pneumonie major outer membrane protein recombinant antigen

目的:为了研究肺炎衣原体(chlamydia pneumoniae,Cpn)抗原作为特异性诊断试剂,本文应用重组技术研制Cpn主要外膜蛋白(major outer membrane protein,MOMP)重组抗原,用于血清IgG抗体的检测。方法:通过构建和表达CpnMOMP重组抗原,应用包涵体纯化法纯化重组抗原后建立ELISA试验,并检测了82份正常人血清及临床标本207份,其中心血管疾病血清118份,呼吸系统疾病血清59份,其他疾病30份。结果:应用克隆技术,获得纯化的Cpn MOMP重组抗原。用重组抗原检测血清IgG,正常人血清阳性检出率仅为3.7%;而207份病人血清,116份为阳性,总阳性率56.0%。其中心血管疾病血清118份,阳性69份,阳性率58.5%;呼吸系统疾病血清59份,阳性38份,阳性率64.4%;其他疾病患者30份,阳性9份,阳性率仅30.0%。结论:与进口ELISA试剂盒相比阳性率基本相符,统计分析无显著意义,但重组MOMP蛋白的抗原特异性较高,且具有抗原制备较为方便和价廉等特点。由于临床标本数量有限,尚需进一步探索MOMP重组抗原作为Cpn感染疾病的辅助诊断。

Objective：To study on an application of Cpn antigen as a special diagnostic tool,recombined Cpn MOMP antigen were produced by recombination technique and used to detect serum IgG.Methods：By establishing and expressing recombined Cpn MOMP antigen,which was purified and used as antigen in ELISA assays.Sera were obtained from 82 healthy volunteers and 207 patients.The patient samples consisted of 118 sera with cardiovascular diseases,59 with diseases of respiratory system and 30 with other diseases.Results：With cloning technique,we obtained depurative recombined antigen of Cpn MOMP.Cpn IgG-positive results were observed in 56.0% of patients and only in 3.7% of healthy subjects.There were 69 positive sera of 118 with cardiovascular diseases,38 positive of 59 with diseases of respiratory system and 9 positive of 30 with other diseases.The positive rates were 58.5%,64.4% and 30.0%.Conclusion：The positive rate of recombined Cpn MOMP antigen is consistent with the ELISA kit.However,due to limit of samples,further studies are needed to the application in clinical accessorial diagnosis.