【摘要】 目的 用TMB(3,3,5,5-四甲基联苯胺)替代OPD(邻苯二胺)作为ELISA实验中的显色剂，以减少OPD对实验操作人员的危害及对环境的污染。方法 依据国家标准ELISA法进行操作，对小鼠肝炎病毒(MHV)抗体ELISA法的OPD和TMB两种显色系统的稳定性、特异性、敏感性及重复性进行比较。结果 TMB系统较OPD系统:TMB比OPD稳定，相对偏差(15.3%)小于OPD(17.7%); TMB系统特异抗原孔P/N值高于OPD特异抗原孔P/N值,差异显著(P<0.05);酶的使用浓度TMB是OPD的0.25倍，检测灵敏度是OPD的2倍;使用TMB,正常抗原和特异抗原孔批内和批间变异系数分别为3.9%、4.7%和13.0%、11.3%;而OPD分别为2.7%、4.5%和11.2%、9.5%。结论 TMB较OPD稳定性、重复性好，敏感性、特异性强，可以替代OPD作为MHV抗体ELISA法的显色剂。更多还原

【Abstract】 Objective In order to reduce the harm of OPD(O-phenylenediamine) to the operators and pollution to environment, TMB(tetramethylbenzidine) was used as a color reagent to replace OPD in ELISA. Methods The operation was carried out on the basis of GB/T14926.50-2001, to compare stability, specificity, sensitivity and repeatability of two color systems in detection of mouse hepatitis virus(MHV) antibody by ELISA. Result The TMB is more stable than OPD, the relative deviation(15.3%) is smaller than OPD(17.7%); the special antigen P/N value of TMB is higher than OPD and the difference is significant(P<0.05); The working concentration of Enzyme of TMB is 0.25 times that of OPD, and the detection sensitivity is twice that of OPD; When using TMB, The inter-assay and the intra-assay average coefficient of variation of normal antigen and special antigen is 3.9%,13.0% and 4.7%, 11.3% while the OPD is 2.7%,11.2% and 4.5%,9.5%. Conclusion TMB is better than OPD in stability,reproduction, sensitivity and specificitys. TMB as a color reagent may replace OPD in detection of mouse hepatitis virus(MHV) antibody by ELISA.更多还原