酶联免疫吸附试验测定儿童体内抗青霉素类IgG抗体

Enzyme linked immunosorbent assay for detection of IgG against penicillins in children

目的：建立一个测定青霉素类反应性ＩｇＧ的酶联免疫吸附方法（ＥＬＩＳＡ）并用该方法观察儿童血清中的特异性抗体．方法：采用免疫印迹、ＥＬＩＳＡ、半抗原抑制试验．结果：青霉素类和牛血清白蛋白结合成复合物，该复合物能作为ＥＬＩＳＡ中的包被抗原（１００ｍｇ·Ｌ－１）．样品稀释倍数和吸光度间有良好的相关性，相关系数０９９１８．运用ＥＬＩＳＡ在１３例临床可疑青霉素过敏的患儿中检测出４例青霉素抗体，１例青霉素和氨苄西林抗体，３例青霉素、氨苄西林、哌拉西林抗体．结论：成功的建立了一个ＥＬＩＳＡ方法，针对青霉素的ＩｇＧ应答在儿童具有异质性．ＩｇＧ抗体不但能识别青霉素分子本身，还能识别降解产物以及来自复合物的新抗原决定族．青霉素、氨苄西林、哌拉西林间有免疫交叉反应．

AIM: To establish an enzyme linked immunosor￣bent assay (ELISA) for detection of IgG against penicillins and then investigate this antibody response in children. METHODS: Western blotting, ELISA, and hapten inhibition assay were used. RESULTS: Penicillins reacted with bovine serum albumin (BSA) to generate conjugate which was used as coating antigen (100 mg·L -1 ) in ELISA. A good correlation was found between absorbance (A) and dilutions of sample with a linear coefficient of 0 9918. Thirteen subjects who were clinically suspicious of penicillin allergy were tested for specific IgG by ELISA, 4 positive for benzylpenicillin, 1 for benzylpenicillin and piperacillin, and 3 for benzylpenicillin, piperacillin and ampicillin. CONCLUSION: An ELISA was successfully established and penicillin reactive IgG response in children was heterogeneous. IgG antibodies recognized benzylpenicillin molecule, degraded products and new antigenic determinant from conjugate. Cross reactions occurred among benzylpenicillin, ampicillin and piperacillin. Inhibitory effects of three opioid receptor agonists on synaptic transmission YUAN Xiao Ru ( Department of Physiology, Nanjing Medical University, Nanjing 210029, China )