Development of the high sensitivity measurements for drug-allergy

药物变态反应采用药物抗原决定簇超敏检测法和阻力淋巴细胞转化试验。用3种不同交联点的头孢克洛/卵清蛋白复合体进行抗体检测。首先，头孢克洛的侧链(R1)被头孢克洛过敏患者的抗头孢克洛抗体识别。相反，IgM抗体与头孢克洛的7-氨基头孢烷酸(7-AMA)反应。竞争酶联免疫吸附试验显示，由α-苯甘氨酸侧链组成的氨苄西林、贝氨西林、头孢克洛和头孢氨苄对抗头孢克洛侧链抗体具有竞争性抑制作用。抗头孢克洛Ig G抗体与α-苯甘氨酸(R1)侧链反应呈剂量反应关系，但不与对氨基苯乙酸和7-AMA反应。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·L，与其他免疫抗体的亲和力基本相同。DLST阳性3例，假阳性3例。而IL-2则在药物刺激后4~8小时内检测到。此外，所有有IL-2产生的患者都检测到了IL-2mRNA，提示IL-2蛋白和mRNA的检测方法比DLST法更敏感，是一种有效的药物过敏诊断方法。

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.