CAJ | 학술논문

目的表达、纯化结核分枝杆菌rESAT-6-CFP-10(rEC)融合蛋白,评价其在结核病血清学诊断方面的应用价值.方法用镍离子螯合亲和层析柱纯化融合蛋白,用ELISA法检测血清样本中的抗结核抗体,以37例PPD阴性患者正常血清A492+2s为正常限值,评价rEC诊断的特异性及灵敏度.结果 rEC融合蛋白在大肠埃希菌中以可溶性非包涵体形式表达,表达浓度为0.19 mg/mL.rFC检测抗结核抗体在PPD阳性健康人群中的特异性为100%(30/30),在痰涂片或细菌培养阳性和两法均阴性的结核病患者中灵敏度分别为75.56%(34/45)和67.3]%(35/52).结论结核分枝杆菌rEC融合蛋白以可溶性形式高效表达,具有较强的免疫反应性和较高的血清学诊断价值.
목적 표체、순화결핵분지간균rESAT-6-CFP-10(rEC)융합단백,평개기재결핵병혈청학진단방면적응용개치.방법 용얼리자오합친화층석주순화융합단백,용ELISA법검측혈청양본중적항결핵항체,이37례PPD음성환자정상혈청A492+2s위정상한치,평개rEC진단적특이성급령민도.결과 rEC융합단백재대장애희균중이가용성비포함체형식표체,표체농도위0.19 mg/mL.rFC검측항결핵항체재PPD양성건강인군중적특이성위100%(30/30),재담도편혹세균배양양성화량법균음성적결핵병환자중령민도분별위75.56%(34/45)화67.3]%(35/52).결론 결핵분지간균rEC융합단백이가용성형식고효표체,구유교강적면역반응성화교고적혈청학진단개치.
Objective To express and purify rESAT-6-CFP-10( rEC) fusion protein of Mycobacterium tuberculosis H37Rv and evaluate its application in tuberculosis serodiagnosis. Methods The rEC fusion protein was purified by nickel-chelate affinity chromatography, and anti-tuberculosis antibody was analyzed by ELISA, the specificity and sensitivity of rEC were evaluated by the positive cut off value determined as A492 + 2s in 37 serum samples from PPD-negative healthy people. Results The rEC fusion protein existed in soluble form in E. coli, and the concentration was 0.19 mg/mL. The specificity in PPD-positive healthy people was 100%(30/30) by anti-tuberculosis antibody detection of rEC, and the sensitivity in smear or germiculture positive and negative tuberculosis patients were 75.56%(34/45) and 67.31%(35/52), respectively. Conclusions The rEC fusion protein can be highly expressed in soluble form in E. coli. It has good immunoreactivity and serodiagnosis value.