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一种用于检测人类和灵长类动物血浆肾素的多量程酶联免疫吸附测定法。

A multirange ELISA for the measurement of plasma renin in humans and primates.

作者信息

Ménard J, Bews J, Heusser C

出版信息

J Hypertens Suppl. 1984 Dec;2(3):S275-8.

PMID:6400374
Abstract

A sensitive solid phase enzyme-linked immunosorbent assay (ELISA) has been developed using two anti-human renin monoclonal antibodies, which were shown to bind both human as well as primate renin at two different epitopic sites. One monoclonal antibody (3-36-16) was used to coat each well of a 96 well microtitre plate in which renin, contained in 5 to 20 ul of plasma, was allowed to react. Plates were then incubated with the gamma globulin (gamma G) fraction of a rabbit anti-human renin serum followed by development with sheetp anti-rabbit gamma G conjugated to alkaline phosphatase. Quantification was carried out by the addition of the alkaline phosphatase substrate, p-nitrophenylphosphate, which produced a colorimetric reaction. The sensitivity of the assay is 25 pg ml-1. The method recognises both active and inactive renin from plasma, kidney, amniotic fluid and chorionic cells. Plasma renin can be measured within 6 hours when values are greater then 150 pg ml-1 or within 24 hours when plasma values are less than 150 pg ml-1. The ELISA has already been used to measure total immunoreactive renin in plasma obtained from patients with several forms of hypertension. The values ranged from 25 pg ml-1 in patients with primary aldosteronism to as large as 60 ng ml-1 in a patient with a renin-secreting tumour.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

利用两种抗人肾素单克隆抗体开发了一种灵敏的固相酶联免疫吸附测定(ELISA)法,这两种抗体可在两个不同的表位位点与人肾素以及灵长类肾素结合。一种单克隆抗体(3 - 36 - 16)用于包被96孔微量滴定板的每孔,使5至20微升血浆中的肾素在其中发生反应。然后将板与兔抗人肾素血清的γ球蛋白(γG)部分孵育,接着用与碱性磷酸酶偶联的抗兔γG进行显色。通过添加碱性磷酸酶底物对硝基苯磷酸酯进行比色反应来进行定量。该测定法的灵敏度为25皮克/毫升。该方法可识别来自血浆、肾脏、羊水和绒毛膜细胞的活性和非活性肾素。当血浆肾素值大于150皮克/毫升时,可在6小时内检测出血浆肾素;当血浆值小于150皮克/毫升时,则可在24小时内检测出。ELISA已用于测量多种高血压患者血浆中的总免疫反应性肾素。其值范围从原发性醛固酮增多症患者的25皮克/毫升到肾素分泌肿瘤患者高达60纳克/毫升不等。(摘要截短至250字)

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