用于检测血清中针对B群脑膜炎奈瑟菌15型调理素的流式细胞术检测法。

Flow cytometric assay for the measurement of serum opsonins to Neisseria meningitidis serogroup B, serotype 15.

作者信息

Sjursen H, Bjerknes R, Halstensen A, Naess A, Sørnes S, Solberg C O

机构信息

Medical Department B, University of Bergen, Haukeland Hospital, Norway.

出版信息

J Immunol Methods. 1989 Jan 17;116(2):235-43. doi: 10.1016/0022-1759(89)90209-3.

DOI:10.1016/0022-1759(89)90209-3

PMID:2492053

Abstract

A flow cytometric phagocytosis assay has been developed for the measurement of human serum opsonins to serogroup B meningococci. Live bacteria and bacteria inactivated by heat, formalin or ethanol were labelled with fluorescein-isothiocyanate (FITC). The bacteria were opsonized with sera from patients with group B meningococcal disease and sera from healthy controls, and phagocytosis determined by combined measurements of FITC-fluorescence and forward angle light scatter. Optimal sensitivity was obtained using viable bacteria, 5% serum, 20 bacteria per leukocyte capable of phagocytosis, 7.5 min opsonization time, 5 min phagocytosis time, 37 degrees C, and continuous agitation during opsonization and phagocytosis. The opsonic activity of sera from convalescent patients was markedly higher than that of sera from patients with acute illness. Only minor day-to-day and interindividual variations were observed. The flow cytometric phagocytosis technique is a rapid and reproducible method for the measurement of serum opsonins to meningococci.

摘要

已开发出一种流式细胞术吞噬试验，用于测定人血清中针对B群脑膜炎球菌的调理素。用异硫氰酸荧光素（FITC）标记活细菌以及经热、福尔马林或乙醇灭活的细菌。用B群脑膜炎球菌病患者的血清和健康对照者的血清对细菌进行调理，通过联合测量FITC荧光和前向角光散射来确定吞噬作用。使用活细菌、5%血清、每白细胞20个具有吞噬能力的细菌、7.5分钟的调理时间、5分钟的吞噬时间、37℃以及在调理和吞噬过程中持续搅拌，可获得最佳灵敏度。恢复期患者血清的调理活性明显高于急性期患者血清。仅观察到轻微的每日和个体间差异。流式细胞术吞噬技术是一种快速且可重复的方法，用于测定血清中针对脑膜炎球菌的调理素。