Kannoujia Dileep Kumar, Nahar Pradip
Institute of Genomics and Integrative Biology, Council of Scientific and Industrial Research, Delhi, India.
Biotechniques. 2009 May;46(6):468-72. doi: 10.2144/000113118.
The enzyme-linked immunosorbent assay (ELISA) technique is the backbone of the diagnostic assay for detection of infectious and allergic diseases. Here, we demonstrate a unique ELISA method for the detection of an antigen or antibody, in which ELISA steps are carried out under pressure instead of conventional thermal incubation. Pressure-mediated ELISA (PELISA), carried out in 1 h shows more than a 2-fold increase in absorbance value than the control experiment carried out at the same time and temperature without applying pressure. Estimation of total IgE by the 1-h PELISA method gives similar absorbance value (1.081+/-0.031, 823.12 IU) to that obtained by 3-h heat-mediated ELISA on an activated surface (HELISA) (1.165+/-0.037, 810.96 IU). Since PELISA is sensitive, specific, and reproducible (intra- and interassay CVs were 6.47% and 9.65%, respectively), it could be an excellent alternative to HELISA or conventional ELISA procedures.
酶联免疫吸附测定(ELISA)技术是用于检测感染性疾病和过敏性疾病的诊断测定的核心技术。在此,我们展示了一种用于检测抗原或抗体的独特ELISA方法,其中ELISA步骤是在压力下而非传统的热孵育条件下进行的。在1小时内完成的压力介导ELISA(PELISA),与在相同时间和温度下未施加压力的对照实验相比,吸光度值增加了两倍多。通过1小时PELISA方法对总IgE的估计,与在活化表面上通过3小时热介导ELISA(HELISA)获得的吸光度值(1.081±0.031,823.12 IU)相似(1.165±0.037,810.96 IU)。由于PELISA具有灵敏、特异且可重复的特点(批内和批间变异系数分别为6.47%和9.65%),它可能是HELISA或传统ELISA方法的极佳替代方法。