Establishment and evaluation of time resolved fluoroimmuno assay for quantification of total immuno globulin E / 重庆医学

ABSTRACT

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 .