疗效评价中酶联免疫吸附试验数据处理的方法学研究

The methodological study of ELISA data processing in efficacy evaluation

目的:对酶联免疫吸附试验(ELISA)数据处理进行方法学研究,为疗效评价提供准确信息。方法:采用梯度稀释法,以化学发光免疫(CL)、时间分辨荧光免疫(TRIF)检测对照,应用ELISA检测技术,对HepG2.2.15细胞株培养上清液中HBsAg和HBeAg进行光密度(OD)值检测,进行数据处理的方法学研究。结果:与CL、TRIF检测结果呈直线相关不同,ELISA检测OD值与病毒浓(滴)度不成直线关系,曲线拟合后可进行未知样的样品病毒浓(滴)度计算,样品冰冻复融对检测结果有较大影响。采用滤光片检测技术的酶标仪其OD值与病毒浓(滴)度呈S线关系,而采用全波长酶标仪其OD值与病毒浓(滴)度呈抛物线关系。结论:ELISA检测数据应曲线拟合进行转换换算,以正确求出样品浓度(含量)。冰存样品和采用滤光片技术检测时建议样品进行合适稀释。

Objective：To perform a methodological study of ELISA data processing in the efficacy evaluation. Methods： With the use of gradient dilution method and comparison with data detected by chemiluminescence immunoassay and time-resolved fluorescence immunoassay, the supernatants of HepG 2. 2. 15 cell lines were detected for HBsAg and HBeAg with method of ELISA, a methodological study of ELISA data processing based on the curve fitting for the efficacy evaluation was carried. Results： Different with the data detected by chemiluminescence immunoassay and time-resolved fluorescence immunoassay which had a linear correlation with gradient dilution , the value of OD of ELISA measurement had no linear correlation with gradient dilution , showed a similar parabolic curve fitting. Sample thawing had a greater impact on the test results. The OD value had a S curve relationship with the sample concentration when using two filter microplate readers produced in home, and parabolic relationship using filter microplate reader, parabolic curve when using two full wavelength microplate readers. Conclusion： The ICso values must be calculated by fitting the inhibition data to a dose-dependent curve. It is suggested that when sample thawing or filter microplate reader are used to detect the appropriate sample dilution should be applied.