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基于多态性膜蛋白 C 的 B 细胞表位开发猪衣原体特异性抗体酶联免疫吸附试验。

Development of a Chlamydia suis-specific antibody enzyme-linked immunosorbent assay based on the use of a B-cell epitope of the polymorphic membrane protein C.

机构信息

Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Gent, Belgium.

Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, MD, USA.

出版信息

Transbound Emerg Dis. 2018 Apr;65(2):e457-e469. doi: 10.1111/tbed.12783. Epub 2018 Jan 4.

Abstract

Chlamydia suis infections lead to economic loss in the pork industry. Chlamydia suis infections could be successfully treated with tetracyclines until the appearance of a tetracycline resistant phenotype, which was acquired via horizontal gene transfer of the tet(C) gene. Given the importance of C. suis as a swine pathogen and as a recently emerged tetracycline resistant pathogen with zoonotic potential, our aim was to develop a sensitive C. suis-specific antibody ELISA based on the polymorphic membrane proteins (Pmps). Chlamydia Pmps are important virulence factors and candidate antigens for serodiagnosis. We identified nine Pmps (PmpA to I) in C. suis strain MD56 using a recently developed Hidden-Markov model. PmpC was the most promising candidate for the development of a C. suis-specific antibody ELISA as the protein was absent in C. abortus, C. pecorum and C. psittaci which also infect pigs and as the protein contained C. suis-specific amino acid regions, absent in C. trachomatis PmpC. We identified an immunodominant B-cell epitope in C. suis PmpC using experimental porcine sera. The sensitivity and specificity of the PmpC ELISA was compared to the complement fixation test (CFT) and to a recombinant MOMP ELISA using experimental sera. The PmpC ELISA detected all positive control sera and was in contrast to CFT and the rMOMP ELISA 100% C. suis specific as positive control sera against other Chlamydia species did not react in the PmpC ELISA. The test was successfully validated using slaughterhouse sera and sera from clinically affected pigs. The PmpC ELISA could assist in diminishing the spread of C. suis infections in the pork industry.

摘要

猪衣原体感染导致养猪业经济损失。四环素类药物曾成功用于治疗猪衣原体感染,但随着 tet(C) 基因的水平转移,出现了四环素耐药表型。鉴于猪衣原体作为猪病原体的重要性,以及作为一种最近出现的具有人畜共患潜力的四环素耐药病原体,我们的目标是开发一种基于多态性膜蛋白(Pmps)的敏感猪衣原体特异性抗体 ELISA。衣原体 Pmps 是重要的毒力因子和血清学诊断候选抗原。我们使用最近开发的隐马尔可夫模型在猪衣原体 MD56 株中鉴定了 9 种 Pmps(PmpA 至 I)。PmpC 是开发猪衣原体特异性抗体 ELISA 的最有前途的候选抗原,因为该蛋白在牛衣原体、绵羊衣原体和鹦鹉热衣原体中不存在,这些衣原体也感染猪,并且该蛋白含有猪衣原体特异性的氨基酸区域,在沙眼衣原体 PmpC 中不存在。我们使用实验性猪血清鉴定了猪衣原体 PmpC 中的一个免疫显性 B 细胞表位。使用实验性血清比较了 PmpC ELISA 的敏感性和特异性与补体结合试验(CFT)和重组 MOMP ELISA。PmpC ELISA 检测了所有阳性对照血清,与 CFT 和 rMOMP ELISA 不同,阳性对照血清针对其他衣原体物种的反应在 PmpC ELISA 中为 100%猪衣原体特异性,因为针对其他衣原体物种的阳性对照血清没有反应。该试验使用屠宰场血清和临床感染猪的血清成功验证。PmpC ELISA 可以帮助减少猪衣原体感染在养猪业中的传播。

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