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生物素-亲和素偶联放大竞争酶联免疫吸附法测定甲氨蝶呤

, PP. 1044-1046

Keywords: 甲氨蝶呤,生物素,亲和素,放大竞争酶联免疫吸附法

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Abstract:

目的将生物素-亲和素偶联放大竞争酶联免疫吸附法(BA-cELISA)引入甲氨蝶呤血药浓度检测,建立一种快速、简便、敏感、特异的甲氨蝶呤检测方法。方法将NH2活性生物素与MTX单抗IgG化学连接,采用亲和素-HRP酶标记系统,建立放大竞争酶联免疫吸附法,连续制备5批试剂盒,对试剂盒进行批内批间精密度、敏感性、特异性、准确性和稳定性进行验证,检测口服MTX的类风湿关节炎患者血清MTX浓度,并对其检测结果进行判定。结果建立的生物素-亲和素偶联放大竞争酶联免疫吸附法测定甲氨蝶呤的方法具有很好的敏感性和特异性,检测MTX的灵敏度达到0.087ng/mL,检测下限为0.137ng/mL,检测线性范围为0.1~10ng/mL;批内RSD为3.3%~8.7%,平均批内差异为6.5%,批间RSD为7.6%~9.9%,重复性教好,检测回收率为94.2%~112.8%。结论生物素-亲和素偶联放大竞争酶联免疫吸附法检测MTX,操作简单,具有较好的灵敏度和特异性。

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