唾液孕酮直接固相化学发光免疫分析

Direct solid-phase chemiluminescence immunoassay for salivary progesterone.

作者信息

De Boever J, Kohen F, Vandekerckhove D

出版信息

Clin Chem. 1986 May;32(5):763-7.

Abstract

A simple, direct chemiluminescence immunoassay for progesterone in mixed, unstimulated saliva is described. We use purified polyclonal anti-progesterone antibodies covalently coupled to polyacrylamide beads and progesterone-11 alpha-hemisuccinyl-aminobutylethyl isoluminol as the chemiluminescent ligand marker. Bound and free ligand are separated by simple centrifugation. The detection limit of the assay is 1.5 pg per tube (38 pmol/L). Intra- and interassay coefficients of variation for low, medium, and high progesterone concentrations are 7.9, 6.8, 8.8% and 9.3, 6.9, 8.5%, respectively. Analytical recovery of added progesterone is 99.6%. Mean +/- SD progesterone concentrations (pmol/L) in saliva are 178 +/- 46 in the follicular phase, 313 +/- 90 in the periovulatory phase, and 658 +/- 166 in the luteal phase of the menstrual cycle. Correlation between progesterone concentrations in serum (assayed by RIA after extraction) and in saliva is good (r = 0.88, p less than 0.001, n = 96). The assay is simple, fast (4 h, including 1.5 h of incubation time), and reliable.

摘要

本文描述了一种用于检测混合、未刺激唾液中孕酮的简单直接化学发光免疫分析法。我们使用与聚丙烯酰胺珠共价偶联的纯化多克隆抗孕酮抗体，以及孕酮-11α-半琥珀酰氨基丁基异鲁米诺作为化学发光配体标记物。通过简单离心分离结合和游离配体。该分析方法的检测限为每管1.5 pg（38 pmol/L）。低、中、高孕酮浓度的批内和批间变异系数分别为7.9%、6.8%、8.8%和9.3%、6.9%、8.5%。添加孕酮的分析回收率为99.6%。月经周期卵泡期、排卵期和黄体期唾液中孕酮浓度的平均值±标准差（pmol/L）分别为178±46、313±90和658±166。血清中孕酮浓度（提取后通过放射免疫分析法测定）与唾液中孕酮浓度之间的相关性良好（r = 0.88，p < 0.001，n = 96）。该分析方法简单、快速（4小时，包括1.5小时孵育时间）且可靠。