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为了建立猴麻疹病毒(MeV)抗体的快速检测方法,本研究采用基因工程技术制备MeV的MeV-32基因原核表达产物即重组MeV-32蛋白作为抗原诊断试剂,以期建立免疫梳方法用于特异性检测实验猴血清中抗MeV的抗体Ig G。结果显示,最佳抗原包被量为0.1 mg/m L;制备好的免疫梳均能够特异性检测到相应的猴MeV阳性血清而不与其他病毒血清间发生交叉反应,表明该检测方法特异性强;敏感性分析结果表明,MeV-32蛋白能够敏感地检测到1∶100倍稀释的MeV阳性血清;稳定性和重复性试验结果表明,同一样品重复检测3次,变异系数(CV)均小于10%;利用该检测方法在对10份可疑猴血清样品进行检测,免疫梳方法与ELISA检测结果一致率为100.0%,Kappa=1.000。上述结果表明,该检测方法灵敏度高、特异性强、重复性好,具有良好的实用性。
In order to establish a rapid detection method for rhesus monkey measles virus (MeV), MeV-32 recombinant MeV-32 protein was used as a diagnostic reagent for antigen in MeV-transgenic mice in order to establish an immune comb method for specificity The anti-MeV antibody Ig G was tested in the serum of experimental monkeys. The results showed that the optimal antigen coating amount was 0.1 mg / m L; the prepared immunized combs could specifically detect the corresponding monkeys MeV-positive sera without cross-reactivity with other virus sera, indicating that the detection method is specific The results of sensitivity analysis showed that the MeV-32 protein could detect the MeV-positive serum diluted 1: 100 times sensitively. The stability and repeatability test results showed that the coefficient of variation (CV) of the same sample was less than 10%. By using this method, 10 samples of suspected monkey serum were detected. The coincidence rate of ELISA and ELISA was 100.0% and Kappa = 1.000. The above results show that the detection method has high sensitivity, specificity, good repeatability and good practicability.