ELISA检测小鼠IgE类抗OA和抗DNP抗体水平

Establishment of an ELISA for Detection of murine anti--OA and anti--DNP specific IgE

本文应用经两种亲和层析法反复纯化吸收的兔抗小鼠IgE(RAME)建立了检测小鼠抗OA及抗DNP特异性IgE的ELISA方法。纯化的RAME经双扩散,免疫电泳,直接法、间接法和双抗体夹心法ELISA鉴定证实为IgE特异性,不与正常小鼠血清、小鼠IgG各亚类及IgM起交叉反应。抗OA特异性IgE ELISA的批内和批间变异系数分别为5.2％和8.8％,抗DNP特异性IgE的批内和批间变异系数分别为5.0％和5.3％。ELISA与PCA的相关性分析显示:两种方法在抗DNP特异性IgE检测时相关性良好,相关系数r=0.9506(P<0.001);两种方法在检测抗OA特异性IgE时相关性较差,免疫后第14、28、35天的相关系数分别为r=0.6665(P>0.05)、r=0.7412(P>0.05)和r=0.8541(P<0.01)。结果提示:ELISA在检测抗OA特异性IgE时比抗DNP特异性IgE时更易受到非IgE类Ig的竞争。

An ELISA for detection of marine anti-OA and anti-DNP specific IgE was developed. RAME was purified with affinity chroma ftography and proved to be IgE monospecific by double diffusion, immunoelectrophoresis, direct ELISA, indirect ELISA and sandwich ELISA. The vatiation coefficients of intra-assay and inter-assay for anti-OA specific IgE were 5.2% and 8.8%, for anti-DNP specific IgE were 5.0% land 5.3% respectively. The correlation coefficient between ELISA and PCA was 0.9506 (p<0.001) for anti-DNP specific IgE;while for anti-OA specific IgE, the corre lation coefficients between ELISA and PCA were 0.6665(P>0.05), 0,7472(P>0.05) and 0.8541(P<0,01) on day 14, 28 and 35 after immunization respectively. The results showed that the detection of anti-OA specific IgE was more easily interfered than that of lanti-DNP specific IgE by competition of non-IgE class Igs.