采用白桂木凝集素-HRP测定唾液SIgA方法的建立

DETERMINING SIgA IN SALIVA BY SANDWICH ENZYME LINKED ASSAY WITH AHL HRP

目的：建立白桂木凝集素－ＨＲＰ夹心法检测唾液中分泌型ＩｇＡ（ＳＩｇＡ）。方法：制备ＡＨＬ－ＨＲＰ作为酶标凝集素，ＡＨＬ为包被物，确定最适包被浓度及ＡＨＬ－ＨＲＰ工作稀释度，夹心酶联检测ＳＩｇＡ，进行检出限、精密度、准确性等实验并制备标准曲线，初步测定７１例正常人唾液ＳＩｇＡ含量。结果：ＡＨＬ－ＨＲＰ夹心法检测ＳＩｇＡ标准曲线线性检测范围５～３３７ｍｇ／Ｌ。本法测定７１例健康人唾液ＳＩｇＡ含量为（５０９．１５±２９６．７５）ｍｇ／Ｌ。其中５份标本批内平均变异７．４％，批间变异１０．５６％，回收率平均９４％。结论：ＡＨＬ－ＨＲＰ夹心法测定ＳＩｇＡ的作用模式为ＡＨＬ→ＳＩｇＡ→ＡＨＬ－ＨＲＰ，可用于唾液中的ＳＩｇＡ含量的测定。方法简便、准确、便于推广。

Objective:AHL HRP Sandwich assay was developed for the measurement of SIgA Methods:AHL HRP was prepared, and optimum concentration of the coating AHL and working dilution degree of AHL HRP was determined for the measure ment of SIgA with Sandwich enzyme linked assay The working rang of the standard curves,precision and accuracy was performed The standard curves were prepared for SIgA Results:The working range of the standard curves were 5￣337mg/L for SIgA respectively The value of SIgA in the saliva of 71 healthy persons was(509 15±296 75) mg/L with the assay Intra assay cv 7 4% Inter assay cv 10 56% Reclamation was 94% Conclusion:The sandwich enzyme linked assay with AHL HRP is simple and precise,and be used in wide range