酶联免疫吸附试验测定血清苯妥英钠浓度

建立酶联免疫吸附实验测定苯妥英钠血药浓度的方法。方法采用碳二亚胺法将化学修饰后的苯妥英钠与人血清白蛋白结合,再与被氧化物酶结合,作为酶标抗原,自制免抗苯妥钠多克隆抗体。结果抗苯妥英钠ＩｇＧ稀释倍数１：２００、酶标记抗原１：１００时,标准曲线最理想;该方法的测定灵敏度为２．８７μｇ／ｍｌ,线性范围２．９－３０μｇ／ｍｌ,批内变异系数３．３％－１０．２％,批间变异系数５．１％－１３．２％,回收率９０％

Establishment of ELISA for measuring phenytoin in serum

OBJECTIVE: To develop a solid phase, competitive enzyme immunoassay for the measurement of serum phenytoin. METHODS: The chemical modified phenytoin combined with human serum albumin and then conjugated with horseradish peroxidase (HRP) to produce the enzyme labeled phenytoin (DPH-HSA-HRP). The anti-phenytoin antibody was prepared in this lab. RESULTS: The working range and sensitivity of this method were 2.9-30 micrograms/ml and 2.87 micrograms/ml respectively. The intra-assay coefficient of variation (CV) was 3.3%-10.2% (n = 15) and inter-assay CV was 5.1%-13.2%. The recovery of this method was 90%-96%. No significant interference was observed with phenobarbital, primidone, carbamazepine, and valproic acid. The new assay method was compared with HPLC method. A linear regression analysis yielded a slope of 1.03, an intercept 1.38 and a correlation coefficient 0.97. CONCLUSIONS: This solid phase enzyme immunoassay for serum phenytoin appears to be simple, precise, and accurate. It may be readily adopted in clinical laboratory for therapeutic monitoring of phenytoin level in serum.