用多粘菌素B单克隆抗体分析法特异性检测b型流感嗜血杆菌脂寡糖

Specific detection of Haemophilus influenzae type b lipooligosaccharide by a polymyxin B monoclonal antibody assay.

作者信息

Mertsola J, Ramilo O, Sáez-Llorens X, Hanson M S, McCracken G H, Hansen E J

机构信息

Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.

出版信息

J Immunol Methods. 1989 Sep 1;122(2):219-26. doi: 10.1016/0022-1759(89)90267-6.

DOI:10.1016/0022-1759(89)90267-6

PMID:2551967

Abstract

A monoclonal antibody (MAb)-based enzyme immunoassay was developed for detection of Haemophilus influenzae type b (Hib) lipooligosaccharides (LOS). The high affinity of polymyxin B for lipid A was used to bind the Hib LOS to microtiter wells. The immobilized LOS was detected with MAbs directed against the oligosaccharide component of Hib endotoxin. Hib LOS concentrations were measured in in vitro samples and in cerebrospinal fluid (CSF) sample obtained from rabbits with experimental Hib meningitis. The sensitivity of the assay was 1 ng LOS/ml sample and the results obtained with this assay correlated significantly with those obtained with the standard Limulus amebocyte lysate assay. This new assay provides a method for specific detection of Hib LOS in CSF samples and in aqueous laboratory fluids. This general methodology should also be useful for experimental research involving specific LPS/LOS molecules.

摘要

开发了一种基于单克隆抗体（MAb）的酶免疫测定法，用于检测b型流感嗜血杆菌（Hib）脂寡糖（LOS）。利用多粘菌素B对脂质A的高亲和力，将Hib LOS结合到微量滴定孔中。用针对Hib内毒素寡糖成分的单克隆抗体检测固定化的LOS。在体外样品和从患有实验性Hib脑膜炎的兔子获得的脑脊液（CSF）样品中测量Hib LOS浓度。该测定法的灵敏度为1 ng LOS/ml样品，并且该测定法获得的结果与标准鲎试剂法获得的结果显著相关。这种新的测定法提供了一种特异性检测脑脊液样品和实验室水性液体中Hib LOS的方法。这种通用方法也应有助于涉及特定脂多糖/脂寡糖分子的实验研究。