The chimeric UreB, FliD and Omp18 proteins for a sensitive and specific diagnosis of Helicobacter pylori infections.

IF 1.3 Q4 MICROBIOLOGY Iranian Journal of Microbiology Pub Date : 2024-06-01 DOI:10.18502/ijm.v16i3.15768

Hassan Seyyedhamzeh, Safar Farajnia, Mohammad Kargar, Behzad Baradaran, Farshid Kafilzadeh

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Abstract

Background and objectives: Helicobacter pylori is known as the main cause of gastrointestinal diseases including gastritis, gastric ulcer and stomach cancer. Serodiagnosis of H. pylori infection is a noninvasive and rapid method but the efficiency of this method is highly dependent to the antigens used. This study evaluated the efficacy of recombinant UreB-Omp18 and FliD for serodiagnosis of H. pylori infection.

Materials and methods: The genes encoding for fliD, ureB, and omp18 was amplified by PCR and cloned into pET-22b and pET-28a vectors. The constructs were expressed in E. coli BL21 and purified by affinity chromatography. The antigenic properties and diagnostic potential of the recombinant proteins were analysed by immunoblotting and ELISA, respectively.

Results: The recombinant UreB-Omp18 and FliD with molecular weights of 48 kDa and 25 kDa were observed on SDS-PAGE and purified by the Ni-NTA column. The ELISA results showed that the sensitivity and specificity of recombinant UreB-Omp18 protein in serodiagnosis of H. pylori infection were 89% and 83%, respectively. Also, the sensitivity and specificity of the recombinant FliD protein were calculated to be 91% and 76%, respectively.

Conclusion: The results indicated that the recombinant UreB-Omp18 and FliD could diagnose H. pylori infection with high sensitivity and specificity.

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嵌合 UreB、FliD 和 Omp18 蛋白用于幽门螺旋杆菌感染的敏感性和特异性诊断。

背景和目的：幽门螺杆菌是胃炎、胃溃疡和胃癌等胃肠道疾病的主要致病菌。幽门螺杆菌感染的血清诊断是一种非侵入性的快速方法，但这种方法的效率与所使用的抗原有很大关系。本研究评估了重组 UreB-Omp18 和 FliD 在幽门螺杆菌感染血清诊断中的有效性：通过 PCR 扩增 fliD、ureB 和 omp18 的编码基因，并将其克隆到 pET-22b 和 pET-28a 载体中。构建体在大肠杆菌 BL21 中表达，并通过亲和层析进行纯化。分别用免疫印迹法和酶联免疫吸附法分析了重组蛋白的抗原性和诊断潜力：结果：重组蛋白 UreB-Omp18 和 FliD 在 SDS-PAGE 上的分子量分别为 48 kDa 和 25 kDa，并经 Ni-NTA 柱纯化。ELISA 结果显示，重组 UreB-Omp18 蛋白在幽门螺杆菌感染血清诊断中的灵敏度和特异性分别为 89% 和 83%。重组 FliD 蛋白的敏感性和特异性分别为 91% 和 76%：结果表明，重组 UreB-Omp18 和 FliD 能以较高的灵敏度和特异性诊断幽门螺杆菌感染。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Iranian Journal of Microbiology MICROBIOLOGY-

CiteScore

2.40

自引率

7.10%

发文量

审稿时长

12 weeks

期刊介绍： The Iranian Journal of Microbiology (IJM) is an international, multi-disciplinary, peer-reviewed journal that provides rapid publication of the most advanced scientific research in the areas of basic and applied research on bacteria and other micro-organisms, including bacteria, viruses, yeasts, fungi, microalgae, and protozoa concerning the development of tools for diagnosis and disease control, epidemiology, antimicrobial agents, clinical microbiology, immunology, Genetics, Genomics and Molecular Biology. Contributions may be in the form of original research papers, review articles, short communications, case reports, technical reports, and letters to the Editor. Research findings must be novel and the original data must be available for review by the Editors, if necessary. Studies that are preliminary, of weak originality or merely descriptive as well as negative results are not appropriate for the journal. Papers considered for publication must be unpublished work (except in an abstract form) that is not under consideration for publication anywhere else, and all co-authors should have agreed to the submission. Manuscripts should be written in English.