重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2014年
33期
4490-4493,4497
,共5页
免疫球蛋白E%过敏反应%铕标记%时间分辨荧光免疫分析法%方法学评价
免疫毬蛋白E%過敏反應%銪標記%時間分辨熒光免疫分析法%方法學評價
면역구단백E%과민반응%유표기%시간분변형광면역분석법%방법학평개
immunoglobulin E%anaphylaxis%europium marker%time resolved fluoroimmunoassay%methodology evaluation
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。
目的：應用時間分辨熒光免疫分析法（TRFIA）建立總免疫毬蛋白E（TIgE）的定量測定方法。方法採用雙抗體夾心法建立TIgE TRFIA ，併對其進行方法學評價。結果建立的 TIgE TRFIA批內、批間變異繫數（CV）分彆為1．59％～1．68％和5．23％～7．33％；檢測低限為0．25 IU/mL ；線性範圍為1．47～1510．00 IU/mL ；其準確性在允許偏差±10％內；迴收率為97．00％～106．75％；交扠反應試驗和榦擾試驗均能滿足檢測要求；檢測TIgE達15000 IU/mL時仍未見 HOOK效應；與歐矇公司的ELISA法檢測40份臨床樣本結果相關性良好（r＝0．9992，P＜0．01），2種方法間預期偏差在允許範圍（±12．5％）內；檢測健康成人樣本TIgE的參攷值為100 IU/mL。結論建立的TIgE TRFIA具有精密度好，靈敏度高，線性範圍寬，準確度高，特異性彊等優點，能滿足臨床需要。
목적：응용시간분변형광면역분석법（TRFIA）건립총면역구단백E（TIgE）적정량측정방법。방법채용쌍항체협심법건립TIgE TRFIA ，병대기진행방법학평개。결과건립적 TIgE TRFIA비내、비간변이계수（CV）분별위1．59％～1．68％화5．23％～7．33％；검측저한위0．25 IU/mL ；선성범위위1．47～1510．00 IU/mL ；기준학성재윤허편차±10％내；회수솔위97．00％～106．75％；교차반응시험화간우시험균능만족검측요구；검측TIgE체15000 IU/mL시잉미견 HOOK효응；여구몽공사적ELISA법검측40빈림상양본결과상관성량호（r＝0．9992，P＜0．01），2충방법간예기편차재윤허범위（±12．5％）내；검측건강성인양본TIgE적삼고치위100 IU/mL。결론건립적TIgE TRFIA구유정밀도호，령민도고，선성범위관，준학도고，특이성강등우점，능만족림상수요。
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 .
