低效价红细胞抗体检测方法ROC曲线分析

Evaluation of four methods on the detection of low titer antibody and optimization of testing conditions

目的应用ROC评价4种低效价抗体的检测方法,通过正交设计优化低效价抗体检测条件。方法分别用盐水法、聚凝胺法、木瓜酶法,微柱凝胶法对216例标本抗体筛查,绘制ROC曲线比较4种检测方法;设计正交试验,优化低效价抗体的检测条件,利用极差分析法寻找最佳检测条件组合。结果盐水法、聚凝胺法、木瓜酶法、微柱凝胶法ROC曲线下面积分别为0.56、0.93、0.90、0.96;正交试验显示,处理筛选细胞的试剂极差最大,为8.0,微柱凝胶法检测低效价抗体评分最高水平分别为反应时间30 min,筛查细胞浓度2%,木瓜酶处理筛选细胞和筛选细胞所含抗原为纯合子。结论微柱凝胶法诊断效率最高,优于其它3种检测方法;该法检测低效价抗体最佳检测条件组合为:反应时间30 min,筛查细胞浓度2%,木瓜酶处理筛选细胞和筛选细胞所含抗原为纯合子。

Objective To evaluate the four methods on the detection of low titer antibody using ROC,and to optimize the testing conditions using orthogonal design. Methods Saline method,Polybrene method,Papaya enzyme method and micro-column gel method were performed respectively to detect 108 specimens with low titer antibody whose titer was less than or equal to 2 and also to detect another 108 negative cases. ROC curve was drawn to compare the four methods. Thereafter,an orthogonal experiment was designed to optimize the testing conditions,so as to obtain the best combination of testing conditions by range analysis. Results The areas under ROC curve of Saline method,Polybrene method,Papaya enzyme method,and micro-column gel method were 0. 56,0. 93,0. 90,and 0. 96,respectively. The scores under 15 min,30min,and60 min by orthogonal experiment with micro-column gel method were 18. 0,20. 0,and 16. 0,respectively. The scores under cell concentrations of 2%,1%,and 0. 5% were 22. 0,20. 0,and 12. 0,respectively. The scores of saline,low ionic strength salt solution,and papaya enzyme were 6. 0,18. 0,and 30. 0,respectively. The scores of antigens found on manufactured cell,homozygous cell,and heterozygous cell were 14. 0,22. 0,and 18. 0,respectively. The largest discrepancy was found in the reagents used to process cells for screening,which ranged up to a maximum of 8. 0. Conclusion Microcolumn gel method was the most efficient method and better than the other three methods. The best combination of testing conditions of micro-column gel method was the following： 30 min of reacting time,2% cell concentration for screening,treatment with papaya enzyme,and selection of cells with homozygous antigens.