胃蛋白酶原Ⅱ时间分辨荧光免疫分析法的建立

Time Resolved Fluoroimmunoassay of Pepsinogen Ⅱ

建立胃蛋白酶原Ⅱ (PGⅡ )的时间分辨荧光免疫分析法 (TRFIA)。包被PGⅡ单抗 810 1# 和使用DTTA络合Eu3 + 标记PGⅡ单抗 810 2 # ,建立PGⅡTRFIA。结果用Log -Logit函数和四参数Logit函数数据处理。批内和批间CV分别为 2 .1%和 3.8% ,平均回收率为 10 4 .6 % ,灵敏度为 0 .0 2 μg/L。标准曲线呈直线 ,ED50 为 16 .2 1μg/L。与PGⅠ、AFP、CA5 0、CA12 5及CA2 4 2等无交叉反应。Eu3 + 标记单抗在 - 30℃保存 ,可达一年以上 ,免疫活性基本无损失。同批试剂连续分析 8个月结果稳定 ,与IRMA测定结果基本吻合。应用TRFIA检测血清PGⅡ具有灵敏度高、操作简便和时间短等特点 ,是一种适用于人群胃病普查的良好方法 ,具有推广应用价值。

A new time resolved fluoroimmunoassay(TRFIA) using a double antibody method for human pepsinogenⅡ (PGⅡ) in serum was developed, with the monoclonal antobody(McAb) 8101 # coated on the 96 well solid phase, binding to a specific antigenic site on the PGⅡ, and the europoum-labelled PGⅡ McAb 8102 # prepared through the chemical reaction with europium-chelate of N-(p-isothiocyanatobenzy1)-diethylenetriamine-N,N,N,N-tetraacetic acid, directing against a different antigenic site on the PGⅡ.The fluorescencl enhancement system was an enhancement solution mainly containing β-naphthoyltrifluoroacetone.The intra-and interassay CV of the TRFIA were 2.1% and 3.8%, and the average recovery rate was 104.6%, the sensitivity was 0.02μg/L.The TRFIA provided a linear dose response with ED 50 of 16.21μg/L.The cross-reaction with PGⅠ,AFP,CA50,CA125 and CA242 was negligible.The europium-labelled 8102 # was stable for more than one year at-30℃ and the results of the TRFIA with the same labelled reagents were stable for eight months, and the results correspond well to IRMA.The method of detecting serum PGⅡ with TRFIA is highly sensitive, less time consumed and easier for operation.