补体结合酶联免疫吸附试验方法的建立

Establishment of Complement Fixation-Enzyme Linked Immunosorbent Assay (CF-ELISA) for the Detection of Brucellosis

关键词:  CF-ELISA  布氏菌病  敏感性  特异性

Key words:  complement fixation-enzyme linked immunosorbent assay (CF-ELISA)  brucellosis  specificity  sensitivity

摘要:

为改进免疫学诊断技术的准确性,研究了一种基于补体结合的免疫学检测新技术——补体结合酶联免疫吸附试验(CF-ELISA)。CF-ELISA技术采用酶标记抗菊糖纯化豚鼠补体C3抗体及其酶显色系统作为补体参与反应的指示系统,用ELISA方法进行补体结合试验。经对布氏菌病抗体检测的初步试验结果显示,CF-ELISA技术可检测到0.01 IU的布氏菌病抗体,灵敏度与间接酶联免疫吸附试验(iELISA)相当,是虎红平板凝集试验(RBPT)试管凝集试验(SAT)的5 000倍、补体结合试验(CFT)的10 000倍。对349份确诊布氏菌病感染群牛、羊血清的检测结果显示,CF-ELISA、iELISA、CFT、SAT、RBPT的阳性率分别为35.82%、36.39%、31.81%、30.09%、36.1%,CF-ELISA与iELISA、CFT、SAT、RBPT的阳性符合率分别为：98.4%、88.8%、80.0%、90.6%。CF-ELISA、iELISA、CFT、SAT、RBPT对490份布氏菌病阴性群牛、羊血清的阴性率分别为100%、99.6%、100%、99.4%、99.8%,CF-ELISA与iELISA、CFT、SAT、RBPT的阴性符合率分别为：99.6%、100%、99.4%、99.8%。研究表明,CF-ELISA是具有高特异性和高敏感性的布氏菌病免疫学检测技术。

Abstract:

A new complement fixation-enzyme linked immunosorbent assay (CF-ELISA) method was established to improve accuracy rate of immunological test. CF-ELISA was test used ELISA mothed,that antibody conjugated Horse Radish Peroxidase (HRP) is anti-complement C3 extracted and purified with inulin from the serum of guinea pigs and the enzyme-substrate system be used detect complement fixation with antibody-antigen complex. The CF-ELISA method could detect 0.01 IU antibody, which had the same sensitivity with iELISA, and about 5000 times higher than that of RBPT、SAT for brucellosis. The positive rate of 349 serum samples from Brucella-infected flocks of goats/sheep and bovines with CF-ELISA, iELISA, CFT, SAT and RBPT method was 35.82%, 36.39%, 31.81%, 30.09% and 36.1%, respectively. The positive coincidence rate of CF-ELISA to iELISA, CFT, SAT, RBPT was 98.4%, 88.8%, 80.0% and 90.6%. The negative rate of 490 samples collected from Brucella-free flocks with CF-ELISA, iELISA, CFT, SAT and RBPT was 100%, 99.6%, 100%, 99.4% and 99.8%, respectively. The negative coincidence rate of CF-ELISA to iELISA, CFT, SAT, RBPT was 99.6%, 100%, 99.4% and 99.8%. The results showed that CF-ELISA was a promising candidate method for the detection of brucellosis with high specificity and sensitivity.