Development of the high sensitivity measurements for drug-allergy

药物过敏用药物抗原决定簇超敏检测法和药物-淋巴细胞转化试验。在ELISA中，使用3种交联位置不同的头孢克洛/OVA复合物进行抗体检测。首先，从头孢克洛过敏患者中获得的抗头孢克洛IgG抗体识别头孢克洛的侧链（R1）。相反，IgM抗体与头孢克洛的7-氨基头孢烷酸（7-AMA）反应。竞争ELISA结果表明，抗头孢克洛侧链抗体可被α-苯甘氨酸侧链所组成的氨苄西林、倍氨苄西林、头孢克洛和头孢氨苄竞争性抑制。抗头孢克洛IgG抗体与α-苯甘氨酸（R1）侧链呈剂量反应，但与对氨基苯乙酸和7-AMA无反应。此外，BIAcore分析表明抗头孢克洛抗体的亲和力为Ka=2。19 × 10 ~（-3）M ~（-1），Kd=2.56 × 10 ~（-4）M ~（-1），与其它免疫抗体的Kd值基本相同。另一方面，25例患者中3例DLST阳性，3例假阳性。相反，IL-2在药物刺激后4 - 8小时检测到。结果表明，IL-2蛋白和mRNA的检测方法比DLST更敏感，是药物过敏的有效诊断试验。

Drug allergy was examined on the supersensitive detection method for the determinant of drug antigen and drag-lymphocyte blastogenesis test. In ELISA, the antibody detection was carried out using 3 kinds of cefaclor/OVA complex of which the cross-link position differed. First, the side-chain(R 1) of cefaclor was recognized by anti-cefaclor IgG antibodies obtained from cefaclor-allergy patient. In contrast, the IgM antibody reacted at 7-aminocephalosporanic acid (7-AMA) of cefaclor. Competitive ELISA shown that anti-cefaclor side-chain antibodies were competitive inhibitied by ampicillin, becampicillin, cefaclor and cephalexin, which were composed from α-phenylglycine side-chain. Anti-cefaclor IgG antibodies reacted to the α-phenylglycine (R1) side- chain in a dose response, but not p-amino phenyl acetic acid and 7-AMA. Furthermore, BIAcore analysis indicated that the affinity of anti-cefaclor antibody was Ka=2. 19 x 10-3M-1and Kd=2.56 x 10-4 M-l , which were almost equal to other the immune antibody. On the other hand, DLST was positive in 3 out of 25 patients, and false positives were 3 patients. In contrast, IL-2 was detected from 4 to 8 hrs after the stimulation by drugs. Moreover,IL-2 mRNA was detected in all patients with the IL-2 production.These results suggested that the detection method of IL-2 protein and mRNA are more super-sensitive than the DLST, and this method is effective diagnosed test for drug-allergy.