Athey D, McNeil C J
Department of Clinical Biochemistry, Medical School, University of Newcastle Upon Tyne, UK.
J Immunol Methods. 1994 Dec 2;176(2):153-62. doi: 10.1016/0022-1759(94)90309-3.
The use of the highly stable, pH insensitive flavoenzyme, reduced nicotinamide adenine dinucleotide oxidase (NADH oxidase) from the thermophilic organism Thermus aquaticus in combination with alcohol dehydrogenase in an amperometric amplified immunoassay for thyrotropin (TSH) is described. NADH oxidase catalyses the oxidation of reduced nicotinamide adenine dinucleotide (NADH) with concomitant two electron reduction of di-oxygen to hydrogen peroxide. Hydrogen peroxide can be detected by oxidation at a platinum electrode poised at +650mV vs. Ag/AgCl. The enzyme amplification system described has advantages over existing amplification techniques in terms of sensitivity, specificity and operational pH dependence. The electrochemical enzyme-amplified assay for TSH was compared with a spectrophotometric enzyme-amplified system and with a non-amplified electrochemical immunoenzymometric TSH assay. The dynamic range of the electrochemical enzyme-amplified TSH immunoassay was 0.2-100 mIU/l, which was four times that of the enzyme-amplified spectrophotometric assay while the detection limits of both techniques were comparable.
本文描述了将来自嗜热生物嗜热水生栖热菌的高度稳定、对pH不敏感的黄素酶——还原型烟酰胺腺嘌呤二核苷酸氧化酶（NADH氧化酶）与乙醇脱氢酶结合，用于促甲状腺激素（TSH）的安培放大免疫测定。NADH氧化酶催化还原型烟酰胺腺嘌呤二核苷酸（NADH）的氧化，同时将二价氧双电子还原为过氧化氢。过氧化氢可通过在相对于Ag/AgCl为+650mV的铂电极上氧化来检测。所描述的酶放大系统在灵敏度、特异性和操作pH依赖性方面优于现有的放大技术。将TSH的电化学酶放大测定与分光光度酶放大系统以及非放大的电化学免疫酶法TSH测定进行了比较。电化学酶放大TSH免疫测定的动态范围为0.2 - 100 mIU/l，是酶放大分光光度测定的四倍，而两种技术的检测限相当。
