总免疫球蛋白E时间分辨荧光免疫分析法的建立与评价

摘要

Objective To establish a method for quantitative detection of total immunoglobulin E(TIgE) by Time resolved Flu‐oroimmunoassay(TRFIA ) .Methods The method for quantitative detection of TIgE by TRFIA was established on the basis of solidphase double sandwich enzyme linked immunosorbent assay(ELISA) .The methodology was evaluated .Results The TIgE TR‐FIA intra assay and inter assay coefficients of variation (CV) were 1 .59% -1 .68% and 5 .23% -7 .33% ,respectively .The lower limit of detection was 0 .25 IU/mL .The linear range was 1 .47-1 510 .00 IU/mL .The accuracy was within the allowable deviation ( ± 10% ) .The recovery rate was 97 .00% -106 .75% .The cross reaction test and interference experiment could meet the testing requirements .The TIgE TRFIA showed no HOOK effect at least up to 15 000 IU/mL TIgE ,compared with EUROIMMUN ELISA ,the correlation coefficient(r) was 0 .999 2(P<0 .01)for 40 blood specimens in the range of 14 .43-518 .81 IU/mL ,and the expected bias was in the range of acceptable bias (± 12 .50% ) .The reference value 100 IU/mL could be used for a normal ,allergy free adult sample TIgE level detected by TRFIA .Conclusion The established TRFIA for TIgE detection meets the demand of clini‐cal application with good precision ,high sensitivity ,wide linear range ,high accuracy ,specificity and other advantages .%目的：应用时间分辨荧光免疫分析法（TRFIA）建立总免疫球蛋白E（TIgE）的定量测定方法。方法采用双抗体夹心法建立TIgE TRFIA ，并对其进行方法学评价。结果建立的 TIgE TRFIA批内、批间变异系数（CV）分别为1．59％～1．68％和5．23％～7．33％；检测低限为0．25 IU/mL ；线性范围为1．47～1510．00 IU/mL ；其准确性在允许偏差±10％内；回收率为97．00％～106．75％；交叉反应试验和干扰试验均能满足检测要求；检测TIgE达15000 IU/mL时仍未见 HOOK效应；与欧蒙公司的ELISA法检测40份临床样本结果相关性良好（r＝0．9992，P＜0．01），2种方法间预期偏差在允许范围（±12．5％）内；检测健康成人样本TIgE的参考值为100 IU/mL。结论建立的TIgE TRFIA具有精密度好，灵敏度高，线性范围宽，准确度高，特异性强等优点，能满足临床需要。