用于吞噬功能的鲁米诺依赖性化学发光微量测定法。

Luminol-dependent chemiluminescence microassay for phagocytic function.

作者信息

Thomas V L, Sanford B A, Driscoll M S, Casto D T, Ramamurthy R S

机构信息

Department of Microbiology, University of Texas Health Science Center, San Antonio 78284.

出版信息

J Immunol Methods. 1988 Jul 22;111(2):227-32. doi: 10.1016/0022-1759(88)90131-7.

DOI:10.1016/0022-1759(88)90131-7

PMID:3397547

Abstract

A luminol-dependent chemiluminescence (CL) microassay was developed to measure phagocytic function of peripheral blood leukocytes. Buffy coats, obtained by centrifugation of only 100 microliter of whole blood, provided an enriched population of polymorphonuclear leukocytes (PMNs). The total reaction mixture, consisting of leukocytes-luminol-inducer (opsonized zymosan), was 450 microliter. Peak CL activity was seen 5 min after addition of inducer at 37 degrees C with cells tested within 60 min after collection. Tests to determine precision and reproducibility of the microassay gave a coefficient of variation of 8.5% and 11%, respectively. There was no significant difference between the mean peak CL values for 20 healthy adult donors compared to 14 premature neonates, however, the newborns' CL activity declined more rapidly; CL activity was severely depressed in cells obtained from a patient with chronic granulomatous disease. Results suggest that this microassay provides a simple, rapid, and reliable test of phagocytic function in cases where the amount of blood available for testing is limited.

摘要

开发了一种基于鲁米诺的化学发光（CL）微量测定法来测量外周血白细胞的吞噬功能。通过仅对100微升全血进行离心获得的血沉棕黄层，提供了丰富的多形核白细胞（PMN）群体。由白细胞-鲁米诺-诱导剂（调理酵母聚糖）组成的总反应混合物为450微升。在37℃添加诱导剂后5分钟观察到峰值CL活性，收集后60分钟内对细胞进行检测。确定该微量测定法精密度和重现性的测试分别给出了8.5%和11%的变异系数。20名健康成年供体的平均峰值CL值与14名早产新生儿相比无显著差异，然而，新生儿的CL活性下降更快；从慢性肉芽肿病患者获得的细胞中CL活性严重降低。结果表明，在可用于检测的血量有限的情况下，这种微量测定法提供了一种简单、快速且可靠的吞噬功能检测方法。