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阿克氏立克次体的蛋白质组学分析提出 44kDa-OMP 作为立克次体痘诊断的潜在生物标志物。

Proteomic analysis of Rickettsia akari proposes a 44 kDa-OMP as a potential biomarker for Rickettsialpox diagnosis.

机构信息

Institute of Virology, Biomedical Research Center of the Slovak Academy of Sciences, Dúbravská cesta 9, 845 05, Bratislava, Slovak Republic.

Department of Biology, Faculty of Science, University of Hradec Kralove, Hradecká 1285, 500 03, Hradec Králové, Czech Republic.

出版信息

BMC Microbiol. 2020 Jul 8;20(1):200. doi: 10.1186/s12866-020-01877-6.

Abstract

BACKGROUND

Rickettsialpox is a febrile illness caused by the mite-borne pathogen Rickettsia akari. Several cases of this disease are reported worldwide annually. Nevertheless, the relationship between the immunogenicity of R. akari and disease development is still poorly understood. Thus, misdiagnosis is frequent. Our study is aiming to identify immunogenic proteins that may improve disease recognition and enhance subsequent treatment. To achieve this goal, two proteomics methodologies were applied, followed by immunoblot confirmation.

RESULTS

Three hundred and sixteen unique proteins were identified in the whole-cell extract of R. akari. The most represented protein groups were found to be those involved in translation, post-translational modifications, energy production, and cell wall development. A significant number of proteins belonged to amino acid transport and intracellular trafficking. Also, some proteins affecting the virulence were detected. In silico analysis of membrane enriched proteins revealed 25 putative outer membrane proteins containing beta-barrel structure and 11 proteins having a secretion signal peptide sequence. Using rabbit and human sera, various immunoreactive proteins were identified from which the 44 kDa uncharacterized protein (A8GP63) has demonstrated a unique detection capability. It positively distinguished the sera of patients with Rickettsialpox from other rickettsiae positive human sera.

CONCLUSION

Our proteomic analysis certainly contributed to the lack of knowledge of R. akari pathogenesis. The result obtained may also serve as a guideline for a more accurate diagnosis of rickettsial diseases. The identified 44 kDa uncharacterized protein can be certainly used as a unique marker of rickettsialpox or as a target molecule for the development of more effective treatment.

摘要

背景

斑点热立克次体病是由螨传播病原体立氏立克次体引起的发热性疾病。全球每年报告数例该病。然而,立氏立克次体的免疫原性与疾病发展之间的关系仍知之甚少。因此,误诊很常见。我们的研究旨在确定可能改善疾病识别并增强后续治疗的免疫原性蛋白。为了实现这一目标,应用了两种蛋白质组学方法,并进行了免疫印迹确认。

结果

在立氏立克次体的全细胞提取物中鉴定出 316 种独特的蛋白质。发现代表性最强的蛋白组是参与翻译、翻译后修饰、能量产生和细胞壁发育的蛋白组。大量的蛋白质属于氨基酸转运和细胞内运输。此外,还检测到一些影响毒力的蛋白质。膜富集蛋白的计算机分析显示 25 种可能的外膜蛋白含有β桶结构,11 种具有分泌信号肽序列。使用兔和人血清,从其中鉴定出各种免疫反应性蛋白,其中 44 kDa 未鉴定蛋白(A8GP63)具有独特的检测能力。它可将斑点热立克次体病患者的血清与其他立克次体阳性的人血清区分开来。

结论

我们的蛋白质组学分析无疑有助于缺乏对立氏立克次体发病机制的了解。获得的结果也可作为更准确诊断立克次体病的指南。鉴定出的 44 kDa 未鉴定蛋白可用作斑点热立克次体病的独特标志物或作为开发更有效的治疗方法的靶分子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0151/7346478/e0c63dd4ca69/12866_2020_1877_Fig1_HTML.jpg

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