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[新生儿21-羟化酶缺乏所致先天性肾上腺皮质增生症的群体筛查。I. 17α-羟孕酮放射免疫测定的微量滤纸法]

[Neonatal mass-screening for congenital adrenal hyperplasia due to 21-hydroxylase deficiency. I. Microfilter paper methods for radioimmunoassay of 17 alpha-hydroxyprogesterone].

作者信息

Shimozawa K

出版信息

Nihon Naibunpi Gakkai Zasshi. 1983 Dec 20;59(12):1845-9. doi: 10.1507/endocrine1927.59.12_1845.

DOI:10.1507/endocrine1927.59.12_1845
PMID:6609091
Abstract

In order to perform neonatal mass-screening for 21-hydroxylase deficiency (21-OHD), two simplified radioimmunoassay (RIA) methods to estimate 17 alpha-hydroxyprogesterone (17-OHP) were devised, using discs 3 mm in diameter cut from filter paper impregnated with whole blood. One was a direct method to use assay buffer eluent as a sample, and the other was an extraction method to use diethyl ether extract. One disc in the direct method and one-fourth to four discs in the extraction method were used. We used [1, 2, 6, 7-3H(N)]-17-OHP as a tracer, anti-17-OHP-3-carboxymethyloxime-BSA serum as an antiserum and saturated ammonium sulfate to separate the bound from free 17-OHP. The specificity of the antiserum was nearly satisfactory, except for an unnegligible cross-reactivity with 17 alpha-hydroxypregnenolone (17-OH-delta 5P). Both methods were practically easy and rapid, and had satisfactory accuracy and precision. A significant correlation (p less than 0.005) was observed between "Disc-17-OHP" values by both methods and plasma 17-OHP concentrations measured by highly specific RIA, although the values obtained by the direct method were significantly higher (p less than 0.005) than plasma 17-OHP concentrations, which presumably resulted from the cross-reactivity of the antiserum with 17-OH-delta 5P-sulfate. "Disc-17-OHP" values of the untreated or poorly controlled patients with 21-OHD were extremely high compared with normal neonates and children. However, as most premature infants also showed high values, the possibility of giving false-positive results had to be considered.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

为了进行21-羟化酶缺乏症(21-OHD)的新生儿群体筛查,设计了两种简化的放射免疫分析(RIA)方法来估算17α-羟孕酮(17-OHP),使用从浸有全血的滤纸上剪下的直径3毫米的圆片。一种是直接法,使用测定缓冲液洗脱液作为样本,另一种是萃取法,使用乙醚提取物。直接法使用一片圆片,萃取法使用四分之一到四片圆片。我们使用[1,2,6,7-³H(N)]-17-OHP作为示踪剂,抗-17-OHP-3-羧甲基肟-BSA血清作为抗血清,并用饱和硫酸铵分离结合态和游离态的17-OHP。抗血清的特异性几乎令人满意,只是与17α-羟孕烯醇酮(17-OH-δ⁵P)有不可忽略的交叉反应。两种方法实际操作都简便快速,准确性和精密度也令人满意。两种方法测得的“圆片-17-OHP”值与通过高特异性RIA测定的血浆17-OHP浓度之间存在显著相关性(p<0.005),尽管直接法获得的值比血浆17-OHP浓度显著更高(p<0.005),这可能是由于抗血清与17-OH-δ⁵P-硫酸盐的交叉反应所致。与正常新生儿和儿童相比,未经治疗或控制不佳的21-OHD患者的“圆片-17-OHP”值极高。然而,由于大多数早产儿也显示出高值,所以必须考虑出现假阳性结果的可能性。(摘要截短于250字)

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