Tsuji A, Maeda M, Arakawa H, Shimizu S, Ikegami T, Sudo Y, Hosoda H, Nambara T
School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
J Steroid Biochem. 1987;27(1-3):33-40. doi: 10.1016/0022-4731(87)90291-3.
Enzyme immunoassays for 17 alpha-hydroxyprogesterone (17-OHP) were developed. Horseradish peroxidase (HRP), glucose oxidase (GOD), invertase (INV) and glucose-6-phosphate dehydrogenase (G6PDH) were used as label enzymes. Double antibody coated beads or tubes were used for separating the bound and free fractions. Antisera used were prepared by using 4-carboxyethylthio-17-OHP and 3-carboxymethyl oxime-17-OHP-bovine serum albumin as immunogens. The bridge heterologous system was more sensitive than other site heterologous and homologous systems. The minimum amounts of 17-OHP detected were 0.25 and 1.0 pg/tube for fluorescence EIAs using HRP and GOD, and 0.1, 10 and 0.1 pg/tube for chemiluminescence EIAs using GOD, INV and G6PDH, respectively. The reproducibility and correlation with RIA were also studied. The present study demonstrates the feasibility of a neonatal screening for congenital adrenal hyperplasia.
开发了用于检测17α-羟孕酮(17-OHP)的酶免疫测定法。使用辣根过氧化物酶(HRP)、葡萄糖氧化酶(GOD)、转化酶(INV)和葡萄糖-6-磷酸脱氢酶(G6PDH)作为标记酶。使用双抗体包被的珠子或试管来分离结合部分和游离部分。所用抗血清是通过使用4-羧乙基硫代-17-OHP和3-羧甲基肟-17-OHP-牛血清白蛋白作为免疫原制备的。桥联异源系统比其他位点异源和同源系统更敏感。使用HRP和GOD的荧光酶免疫测定法检测到的17-OHP的最低量分别为0.25和1.0 pg/管,使用GOD、INV和G6PDH的化学发光酶免疫测定法检测到的最低量分别为0.1、10和0.1 pg/管。还研究了其重现性以及与放射免疫分析(RIA)的相关性。本研究证明了新生儿先天性肾上腺皮质增生症筛查的可行性。