TRFIA法检测总免疫球蛋白E的分析灵敏度及可报告范围的建立

Establishment of analysis sensitivity and reportable range of total immunoglobulin E detected by time-resolved fluoroimmunoassay

目的探讨时间分辨荧光免疫分析法(TRFIA)检测总免疫球蛋白E(总IgE)的分析灵敏度,并建立可报告范围。方法以试剂盒0 IU/mL的校准品A为空白样本,用校准品A将12 IU/mL的校准品稀释成低浓度系列样本作为分析灵敏度试验样品,空白样本批内重复检测20次,分析灵敏度样本天间重复检测20次,记录每次测定的发光值CPs,计算检测低限(LLD)、生物检测限(BLD)和功能灵敏度(FS)。参考EP6-A文件和相关文献,选用1.21 IU/mL和2 001.01 IU/mL的样本按比例精确配制成不同浓度的系列分析测量范围(AMR)试验样品,重复检测4次,将实测值与预期值作比较,建立该系统的AMR,并选用3份接近AMR上限的高浓度样本,用稀释液做不同倍数稀释后检测,计算稀释回收率,确定该系统最大允许稀释倍数,并结合FS建立临床可报告范围(CRR)。结果本研究总IgE TRFIA系统的LLD可达0.21 IU/mL,BLD可达1.00 IU/mL,FS为1.00 IU/mL,AMR为1.12~1 601.03 IU/mL。确定了可最大允许稀释倍数为1:100,CRR为1.00 IU/mL^160 103 IU/mL。结论本研究总IgE TRFIA系统的灵敏度高,AMR和CRR可满足临床需要。

Objective To establish the analysis sensitivity and reportable range of total immunoglobulin E （TIgE） by time-resolved fluoroimmunoassay （TRFIA）.Methods Calibrator A （0 IU/mL） of TIgE was used as a blank sample.The blank sample was used to dilute the calibrator with low concentration of TIgE （12 IU/mL） to prepare a series of low-concentration samples which would be taken as the experiment samples of analysis sensitivity.Blank samples and other samples were repeatedly in-batch or between-batch detected for 20 times,respectively.Light detection values （CPs） of each detection were recorded,and lower limit of detection （LLD）,biologic limit of detection （BLD） and functional sensitivity （FS） were calculated.EP6-A evaluation protocols and pertinent literature served as the references,a series of analytical measurement range （AMR） samples were prepared with samples at 1.21 IU/mL and 2001.01 IU/mL,then they were detected 4 times each.After we compared the measured value with the expected value,the AMR was established.Three samples,at high concentration level and within the AMR,were diluted with different times of dilution,and then the dilution recovery rate was calculated to determine the max allowable dilution rate.After that,the clinical reportable range （CRR） was established by combining FS.Results In this study,the total IgE TRFIA LLD was up to 0.21 IU/mL,BLD up to 1.00 IU/mL,FS to 1.00 IU/mL,and AMR was 1.12-1 601.03 IU/mL.The maximum allowable dilution ratio was 1：100,CRR was 1.00-160 103 IU/mL.Condusion The TIgE TRFIA system meets the demand of clinical application with high sensitivity,wide AMR and CRR.