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评估一种已开发的种特异性单克隆抗体,用于检测女性患者宫颈内膜标本中的沙眼衣原体感染。

Evaluation of a developed species-specific monoclonal antibody for detecting Chlamydia trachomatis infections in endocervical specimens from female patients.

作者信息

Kumar Amit, Singh S, Salhan S, Mittal Aruna

机构信息

Institute of Pathology (ICMR), Department of Gynecology, Safdarjang Hospital Campus, New Delhi, India.

出版信息

Hybridoma (Larchmt). 2007 Oct;26(5):333-7. doi: 10.1089/hyb.2007.0515.

Abstract

To increase the sensitivity and reliability of immunodiagnostic assay for direct detection of chlamydial genital infection, we developed monoclonal antibodies (MAbs) that recognize the major outer membrane protein (MOMP) of all serovars of Chlamydia trachomatis. ELISA and dot-ELISA were used to select hybridomas that produced anti-C. trachomatis MAbs. Four MAb clones--A12.6, B serogroup specific; C1.5, C serogroup specific; G10.2, species specific; and E9.1, genus specific--were characterized by SDS-PAGE and immunoblot and were reactive with MOMP of C. trachomatis. The species-specific MAb G10.2 had a molecular mass of 40 kDa. The reactivity of developed species-specific monoclonal antibody for detection of C. trachomatis antigen in endocervical specimens was compared with the commercially available DFA test. We found that developed antibody had high sensitivity (97.22%) compared to DFA for detection of chlamydial infection in endocervical samples. It can be used as a reliable method in laboratories for the diagnosis of chlamydial infections.

摘要

为提高直接检测衣原体性生殖器感染的免疫诊断检测的灵敏度和可靠性,我们制备了可识别沙眼衣原体所有血清型主要外膜蛋白(MOMP)的单克隆抗体(MAb)。采用酶联免疫吸附测定(ELISA)和斑点ELISA筛选产生抗沙眼衣原体MAb的杂交瘤。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)和免疫印迹对4个MAb克隆进行了鉴定,分别为:A12.6,B血清群特异性;C1.5,C血清群特异性;G10.2,种特异性;E9.1,属特异性,它们均与沙眼衣原体的MOMP发生反应。种特异性MAb G10.2的分子量为40 kDa。将所制备的种特异性单克隆抗体检测宫颈标本中沙眼衣原体抗原的反应性与市售直接荧光抗体(DFA)检测进行了比较。我们发现,与DFA检测宫颈样本中的衣原体感染相比,所制备的抗体具有较高的灵敏度(97.22%)。它可作为实验室诊断衣原体感染的可靠方法。

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