应用重组致密颗粒抗原6检测诊断弓形虫病的研究  

Application of the recombinant GRA6 in the diagnosis of toxoplasmosis

作　　者：朱翔[1] 钮志林 路文明[1] 丁宁玲 吴燕[1] 王锋[1] 叶建中[1] 沙莉[1] 李扬[1] 龚婵聪 黄金龙[1] 高胜兰 

机构地区：[1]苏州大学附属传染病院肝病科,江苏苏州215007 [2]苏州市吴江区第一医院感染科,江苏苏州215200

出　　处：《中华医院感染学杂志》2015年第24期5530-5533,共4页Chinese Journal of Nosocomiology

基　　金：江苏省科学技术厅社会发展基金资助项目(DS2005027)

摘　　要：目的以纯化的重组致密颗粒抗原6作为检测抗原,建立检测弓形虫IgM和IgG抗体的ELISA新方法,为临床治疗提供参考依据。方法 2007年1月-2011年12月对59份弓形虫阳性血清标本进行检测,并与进口弓形虫ELISA-IgM和IgG试剂盒进行比较,数据采用SPSS 13.0软件进行统计分析。结果 ELISA法优化检测条件为包被抗原浓度为40μg/ml;敏感度比较表明血清稀释度在1∶10~1∶80为优;特异性试验表明IgM阳性的抑制率为97.36%、IgG阳性的抑制率为97.98%;用rGRA6-IgM-ELISA对混合弓形虫IgM阳性和阴性血清的精密度检测表明,IgM阳性的变异系数（CV值）为2.76%,IgM阴性混合血清的CV值为0.45%;用rGRA6-IgG-ELISA对混合弓形虫IgG阳性和阴性血清的精密度检测表明,IgG阳性的变异系数（CV值）为2.89%,IgG阴性混合血清的CV值为1.65%;rGRA6-IgM-ELISA与进口试剂盒的总符合率为91.01%,rGRA6-IgG-ELISA与进口试剂盒的总符合率为94.59%。结论重组抗原rGRA6能被弓形虫感染患者血清IgM和IgG抗体所识别,用重组抗原rGRA6构建的试剂盒诊断弓形虫病具有较高的特异性、敏感性。OBJECTIVE To establish a new method by using IgM-ELISA and IgG-ELlSA,with the purified recombi nant antigen rGRA6 for immunodiagnosis of toxoplasmosis so as to provide references for clinical treatments. METHODS From Jan. 2007 to Dec. 2011, 59 toxoplasmosis positive sera were assayed with this new method. The results were analyzed and compared with the imported TOXO IgM-ELISA and TOXO ]gC,--ELISA kits. SPSS 13.0 was adopted for Statistical analysis. RESULTS The optimized detecting condition was that the coated antigen was 40 μg/ml. Sensitivity comparison showed that the appropriate serum dilutions were in 1 ： 10 -1 ： 80 range. Specificity showed that the IgM positive inhibition ratewas 97.36 % and the IgG positive inhibition rate was 97.98 % The rGRA6-IgM-ELISA showed that the cgefficient of variation （CV） values of IgM-positive and IgM negative pooled sera were 2.76% and 0.45 % respectively. The rGRA6-IgG-ELISA showed that the coefficient of variation （CV） values of IgG-positive and IgG-negative. pooled sera were 2.89% and 1.65%respectively. The total correspondence rate was 91.01 % compared with the imported TOXO lgM-ELISA kit and 94.59% compared with the imported TOXO IgG-EI.ISA kit respectively. CONCLUSION The recombinant antigen rGRA6 can be recognized by toxoplasmosis-infective IgM and IgG in human sera. The kits constructed by the recombinant antigen rGRA6 were demonstrated to be highly sensitive and specific for detection of T. gondii infection.

关 键 词：弓形虫 重组致密颗粒抗原6 酶联免疫吸附试验 

分 类 号：R446[医药卫生—诊断学]