摘要
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。
To research a rapid assay for the detection of antibodies against simian measles virus（MeV）,in this experiment the immuno combarray（IC） was developed using MeV recombinant MeV-32 protein as antigen diagnostic reagents.In result,the optimum coating amount of the antigen was 0.1 mg/m L.The prepared IC was able to specifically detect the positive sera against MeV,and there were no cross reactions with the other viruses.The MeV-32 protein was able to sensitively detect 1 ∶ 100 times MeV positive serum dilution.The same sample was repeated 3 times,and the coefficient of variation（CV） was less than 10%.In the detection of 10 suspected monkey serum samples using the immune detection method,a consistent rate of the comb with the ELISA test was 100%,and Kappa was 1.000.The above-mentioned results showedthat the developed immuno combarray had high sensitivity,strong specificity,good repeatability and practicability.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第11期1341-1348,共8页
Chinese Veterinary Science
基金
国家质检总局科技项目(2013IK031
2015IK089)
福建省科技计划重点项目(2014N0001)
海南省应用技术研究与开发专项项目(ZDXM20130025)
广东检验检疫局科技计划项目(2011GDK44
2013GDK04)
