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动弯杆菌属单克隆抗体的制备与特性分析

Production and characterization of monoclonal antibodies to Mobiluncus species.

作者信息

Fohn M J, Lukehart S A, Hillier S L

机构信息

Department of Medicine, University of Washington, Seattle 98195.

出版信息

J Clin Microbiol. 1988 Dec;26(12):2598-603. doi: 10.1128/jcm.26.12.2598-2603.1988.

Abstract

Members of the genus Mobiluncus are anaerobic motile curved rods which are associated with bacterial vaginosis (BV). Murine monoclonal antibodies (MAbs) to the ATCC type strains of M. curtisii subsp. curtisii, M. curtisii subsp. holmesii, and M. mulieris were produced and characterized by enzyme-linked immunosorbent assay and indirect immunofluorescence assay. Four MAbs were subspecies specific and reacted with M. curtisii subsp. curtisii but not with M. curtisii subsp. holmesii; four were specific for M. mulieris. The remaining antibodies demonstrated some cross-reactivity: three were species specific and reacted with both subspecies of M. curtisii, and one defined an epitope shared by M. curtisii subsp. holmesii and M. mulieris but not by M. curtisii subsp. curtisii. None of the MAbs reacted with a panel of other bacteria commonly present in the vaginas of normal women or women with BV. Examination of the molecular specificities of the antibodies by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting revealed four antibodies which were specific for an 82,000-dalton molecule of M. curtisii subsp. curtisii and five antibodies which bound a major band of M. mulieris at 93,000 daltons. Selected MAbs reacted in the indirect immunofluorescence assay with 24 of 25 Mobiluncus spp. clinical isolates from local women with BV and could be used for direct detection of Mobiluncus spp. in vaginal fluid from a patient with BV.

摘要

动弯杆菌属的成员是厌氧的、可运动的弯曲杆菌,与细菌性阴道病(BV)相关。制备了针对柯氏动弯杆菌亚种柯氏亚种、柯氏动弯杆菌亚种霍氏亚种和女性动弯杆菌ATCC型菌株的鼠单克隆抗体(MAb),并通过酶联免疫吸附测定和间接免疫荧光测定对其进行了表征。四种单克隆抗体具有亚种特异性,与柯氏动弯杆菌亚种柯氏亚种反应,但不与柯氏动弯杆菌亚种霍氏亚种反应;四种对女性动弯杆菌具有特异性。其余抗体表现出一些交叉反应性:三种具有种特异性,与柯氏动弯杆菌的两个亚种都反应,一种确定了柯氏动弯杆菌亚种霍氏亚种和女性动弯杆菌共有的一个表位,但柯氏动弯杆菌亚种柯氏亚种没有。没有一种单克隆抗体与正常女性或患有细菌性阴道病的女性阴道中常见的一组其他细菌发生反应。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹检查抗体的分子特异性,发现四种抗体对柯氏动弯杆菌亚种柯氏亚种的82,000道尔顿分子具有特异性,五种抗体与女性动弯杆菌93,000道尔顿的一条主要条带结合。选定的单克隆抗体在间接免疫荧光测定中与来自当地患有细菌性阴道病的女性的25株动弯杆菌临床分离株中的24株发生反应,可用于直接检测患有细菌性阴道病患者阴道液中的动弯杆菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/16b0/266954/6f1274d8dfdf/jcm00084-0149-a.jpg

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