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获取基于沙眼衣原体重组蛋白的 ELISA 检测方法。

Obtaining an ELISA test based on a recombinant protein of Chlamydia trachomatis.

机构信息

Department of Microbiology, Escuela Nacional de Ciencias Biológicas del Instituto Politécnico Nacional, Carpio y Pro. Plan de Ayala, Col. Casco de Sto. Thomas, C.P. 11340, Mexico City, México.

Virology Laboratory, National Institute of Perinatology, Isidro Espinoza de los Reyes, Montes Urales 800, Col. Lomas Virreyes, CP 11000, Mexico City, Mexico.

出版信息

Int Microbiol. 2019 Dec;22(4):471-478. doi: 10.1007/s10123-019-00074-4. Epub 2019 Apr 11.

Abstract

Chlamydia trachomatis is considered as a public health problem due to its high prevalence and increased rates of gynecological disorders. The major outer membrane protein (MOMP) of this bacterium is the most abundant protein in its membrane and has been evaluated not only as a vaccine development candidate but also is used in many diagnostic tests. The MOMP weighs 69 kDa and contains four variable segments (VS 1-4) separated by constant regions. Several research groups have developed recombinant single-variable segments of MOMP expressed in Escherichia coli cytoplasm. But, all variable segments have been used minimally for the diagnosis of a chlamydial infection. In this experiment, the authors obtained the recombinant MOMP of C. trachomatis (rMOMP) in E. coli rMOMP and extracted, purified, and partially characterized it. This was later used to identify anti-Chlamydia trachomatis antibodies in sera of infertile patients by immunodetection assays, enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence tests. The ELISA test showed high sensitivity and low specificity of 100 and 58.3%, respectively. The above results obtained were linked to the cross-reactivity of antibodies against C. pneumoniae or C. psittaci. Hence, an evaluation was performed to obtain an optimized test for the diagnosis of C. trachomatis infection.

摘要

沙眼衣原体被认为是一个公共卫生问题,因为它的高流行率和增加的妇科疾病发病率。这种细菌的主要外膜蛋白(MOMP)是其膜中最丰富的蛋白质,不仅被评估为疫苗开发的候选物,而且还被用于许多诊断测试。MOMP 重 69kDa,包含四个可变片段(VS1-4),由恒定区隔开。几个研究小组已经在大肠杆菌细胞质中表达了重组单变量 MOMP。但是,所有的可变片段都被最小化地用于诊断衣原体感染。在这个实验中,作者在大肠杆菌 rMOMP 中获得了重组沙眼衣原体 MOMP(rMOMP),并对其进行了提取、纯化和部分特性分析。随后,它被用于通过免疫检测分析、酶联免疫吸附试验(ELISA)和间接免疫荧光试验来鉴定不孕患者血清中的抗沙眼衣原体抗体。ELISA 试验显示出 100%的高灵敏度和 58.3%的低特异性。上述结果与针对肺炎衣原体或鹦鹉热衣原体的抗体的交叉反应性有关。因此,进行了一项评估,以获得一种优化的用于诊断沙眼衣原体感染的测试。

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