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人血清中类胰蛋白酶物质的放射免疫测定

Radioimmunoassay of trypsin-like substance in human serum.

作者信息

Malvano R, Marchisio M, Massaglia A, Giacosa P A, Zannino M, Andriulli A, Burlina A

出版信息

Scand J Gastroenterol Suppl. 1980;62:3-10.

PMID:6935753
Abstract

A radioimmunological method for circulating trypsin-like substances was set up, using cathodic trypsin (isolated from human pancreas) as immunogen, reference standard and material to label, and bovine serum to stabilize the system. The assay scheme included a bound-free separation by polyethylene-glycol precipitation after a 1-h incubation at room temperature. Some relevant points emerged from the validation experiments: a) the assay proved simple and reliable, meeting both requirements of a prompt clinical response and of a high sensitivity level (1.5 ng/ml or less); b) evidence was added to the view that cathodic trypsinogen is the immunoassayable species present in the blood stream; c) the parallelism of the radioimmunological responses of cathodic trypsin and trypsinogen demonstrated that a self-consistent analytical information may be derived using either references; d) the absence of detectable amounts of trypsin-like substances in sera from totally-pancreatectomized patients (3 cases) indirectly confirmed the organ-specificity of the measurement; e) a "normal" level of 27 +/- 8 ng trypsin equivalent/ml (X +/- SD, n = 82) resulted: some much higher literature data, all related to a single radioimmunoassay system (RIA-Gnost trypsin kit, Hoechst, FRG), are seemingly explained by the different purity observed for the reference materials.

摘要

建立了一种用于检测循环中类胰蛋白酶物质的放射免疫方法,使用阴极胰蛋白酶(从人胰腺分离)作为免疫原、参考标准品和标记物,并使用牛血清来稳定系统。检测方案包括在室温下孵育1小时后通过聚乙二醇沉淀进行结合-游离分离。验证实验得出了一些相关要点:a)该检测方法简单可靠,满足了临床快速反应和高灵敏度水平(1.5 ng/ml或更低)的要求;b)进一步证明了阴极胰蛋白酶原是血液中可通过免疫测定的物质;c)阴极胰蛋白酶和胰蛋白酶原的放射免疫反应的平行性表明,使用任何一种参考物都可获得自洽的分析信息;d)全胰切除患者(3例)血清中未检测到类胰蛋白酶物质,间接证实了该测量的器官特异性;e)得出“正常”水平为27±8 ng胰蛋白酶当量/ml(X±SD,n = 82):文献中一些高得多的数据,均与单一放射免疫分析系统(RIA-Gnost胰蛋白酶试剂盒,德国赫斯特公司)相关,似乎可以用参考物质的不同纯度来解释。

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