期刊论文

Antibody titre quantitation method by ELISA without standard substance based on kinetics of enzyme-catalyzed reactions

中文

重庆医学

1671-8348

2015

44

12

1662-1663,1666

10.3969/j.issn.1671-8348.2015.12.026

湖北省教育厅科研计划资助项目（Q20121808）.

本校为第一机构

目的 建立无标准品的酶联免疫吸附测定(ELISA)抗体滴度定量方法.方法 对血清抗体ELISA检测设置非特异、阴性对照、特异和总抗体检测组.显色早期的时间-吸光度数据经线性拟合的斜率即吸光度变化速度 ,分别记为ν0 、νC 、νS 、νT .基于底物过量时ν值与待测抗体浓度(C)呈直线关系 ,设计ν值函数计算待测样本与阴性对照样本的抗体浓度倍比.以鱼胶原蛋白免疫昆明小鼠的血清特异Ig G抗体检测进行实例分析.结果 函数C/CC = (ν-ν0 )/(νC -ν0 )可计算待测样本与阴性对照样本的特异抗体浓度倍比(滴度).结论 该动力学ELISA抗体滴度检测方法适用于无标准品半定量分析.

Objective To establish a antibody titre quantitation method by ELISA without standard substance. Methods The test groups of non-specific,negative control,specific and total antibodies were set for detecting sera antibodies by ELISA. After linear fitting of the time-absorbance data of early developing, the fitted slopes were used as the velocity of absorbance changing, which were denoted by v0 ,vc ,vs ,vr. Based on that the v-value and the concentration （C） of determinand were linear relationship while the substrate was excessive,the function ...