用于感染的灵敏且特异诊断的嵌合尿素酶B、鞭毛蛋白D和外膜蛋白18蛋白。

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

作者信息

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

机构信息

Department of Microbiology, Faculty of Basic Sciences, Jahrom Branch, Islamic Azad University, Jahrom, Iran.

Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Iran J Microbiol. 2024 Jun;16(3):357-365. doi: 10.18502/ijm.v16i3.15768.

DOI:10.18502/ijm.v16i3.15768

PMID:39005599

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11245343/

Abstract

BACKGROUND AND OBJECTIVES

is known as the main cause of gastrointestinal diseases including gastritis, gastric ulcer and stomach cancer. Serodiagnosis of 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 infection.

MATERIALS AND METHODS

The genes encoding for and was amplified by PCR and cloned into pET-22b and pET-28a vectors. The constructs were expressed in 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 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 infection with high sensitivity and specificity.

摘要

背景与目的

幽门螺杆菌是胃炎、胃溃疡和胃癌等胃肠道疾病的主要病因。幽门螺杆菌感染的血清学诊断是一种非侵入性快速方法，但该方法的效率高度依赖所用抗原。本研究评估重组脲酶B-外膜蛋白18（UreB-Omp18）和鞭毛蛋白D（FliD）用于幽门螺杆菌感染血清学诊断的效能。

材料与方法

通过聚合酶链反应（PCR）扩增编码幽门螺杆菌和的基因，并克隆到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纯化。分别通过免疫印迹和酶联免疫吸附测定（ELISA）分析重组蛋白的抗原特性和诊断潜力。

结果

在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）上观察到分子量分别为48 kDa和25 kDa的重组UreB-Omp18和FliD，并通过镍-亚氨基三乙酸（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学诊断中的敏感性和特异性分别为89%和83%。此外，重组FliD蛋白的敏感性和特异性经计算分别为91%和76%。