巨细胞病毒IgG定量ELISA检测方法的建立及初步应用

Development and preliminary application of quantitative ELISA method for cytomegalovirus IgG

目的建立巨细胞病毒(Cytomegalovirus,CMV)IgG定量ELISA检测方法,并进行初步应用。方法用CMV IgG(50 U/ml)标准品建立CMV IgG ELISA定量检测方法,确定该方法的最佳线性范围;制备室内质控血清并进行标定。对该方法进行重复性、中间精密性、准确性、特异性验证,并与Dia.Pro CMV IgG ELISA试剂盒进行比较。用建立的ELISA法检测689人份健康献浆员血浆的CMV IgG抗体效价。结果建立的CMV IgG定量ELISA检测方法的最佳线性范围为0.000 78～0.05 U/ml;制备的室内质控血清的抗体效价为(5.67±0.55)U/ml;重复性试验检测结果的变异系数为4.6%～9.8%,中间精密性试验检测结果的变异系数为9.7%～10.6%;该方法检测3个浓度标准品的回收率为107.01%～112.97%;样品稀释液和血清中的其他一些干扰因素对检测结果影响较小;该方法与Dia.Pro CMV IgG ELISA试剂盒检测血浆样品的相关系数r=0.81。该方法检测689人份健康献浆员血浆样品的CMV IgG抗体效价≥10 U/ml的占30.8%。结论建立的CMV IgG定量ELISA检测方法操作简便,精密性良好,特异性强,可用于人血浆中CMV IgG的定量检测。

Objective To develop and preliminarily apply a quantitative ELISA method for cytomegalovirus （CMV） IgG. Methods A quantitative ELISA method for CMV IgG was developed by using CMV IgG standard, of which the optimal linear range was determined. Internal quality control serum was prepared and calibrated, based on which the developed method was verified for reproducibility, intermediate precision, accuracy and specificity, and the results were compared with those of Dia. Pro CMV IgG ELISA kit, The CMV IgG titers in plasma samples from 689 donors were determined by the developed method. Results The optimal linear range of developed ELISA method was 0. 000 78 - 0. 05 U / ml, and the antibody titer of prepared internal quality control serum was （5.67 ± 0. 55） U/ml. The coefficient of variation （CV） of result of reproducibility test was 4. 6% - 9. 8%, while that of intermediate precision test was 9. 7% - 10. 6%. The recovery rates of standard at three concentrations determined by the developed method were 107.01% - 112. 97%. The diluent of samples and other interfering factors in serum showed no significant influence on test result. The correlation coefficient （r） of test results by the developed method and by Dia. Pro CMV IgG ELISA kit was 0. 81. The CMV IgG titers in 30. 8% of 689 samples determined by the developed method were not less than 10 U/ml. Conclusion The developed quantitative ELISA method was easy to handle, which showed high precision and specificity and might be used for determination of CMV IgG in human plasma.