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Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology

BACKGROUND AND OBJECTIVES: The detection of Ureaplasma urealyticum is usually done through culture. With the change of the smallest effective factor in culture, we face the lack of growth of these bacteria, which is one of the important reasons to find a suitable alternative for the diagnosis of thi...

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Autores principales: Darvishi, Faleh, Ganji, Ali, Khansarinejad, Behzad, Sadoogh Abbasian, Shabnam, Abtahi, Hamid
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Tehran University of Medical Sciences 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9867610/
https://www.ncbi.nlm.nih.gov/pubmed/36721453
http://dx.doi.org/10.18502/ijm.v14i6.11255
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author Darvishi, Faleh
Ganji, Ali
Khansarinejad, Behzad
Sadoogh Abbasian, Shabnam
Abtahi, Hamid
author_facet Darvishi, Faleh
Ganji, Ali
Khansarinejad, Behzad
Sadoogh Abbasian, Shabnam
Abtahi, Hamid
author_sort Darvishi, Faleh
collection PubMed
description BACKGROUND AND OBJECTIVES: The detection of Ureaplasma urealyticum is usually done through culture. With the change of the smallest effective factor in culture, we face the lack of growth of these bacteria, which is one of the important reasons to find a suitable alternative for the diagnosis of this bacterium. UreD is a protected gene in this bacterium. The aim of this was to evaluate the ability of antigenic regions of UreD protein to bind to patients’ serum antibodies. MATERIALS AND METHODS: Antigenic regions of UreD protein were predicted using IEDB software with five different methods: Emini Surface Accessibility Prediction, Kolaskar and Tongaonkar Antigenicity, Chou and Fasman beta turn prediction, Karplus and Schulz flexibility scale, Ellipro–Epitope prediction based upon structural protrusion. Antigenic regions of UreD gene was clonned, expressed and purified. The antigenicity of this recombinant protein against the antibodies in the serum of people infected with U. urealyticum infections was checked in western blotting. RESULTS: The results showed that the antigenic regions of the UreD protein was producted and its antigenicity was demonstrated in western blotting. Moreover, all sera from patients infected with U. urealyticum reacted to the recombinant antigen. CONCLUSION: Specimens from people infected with U. urealyticum infection was positive in Western blotting suggesting that UreD protein has antigenic properties. Therefore, it can be used as a suitable candidate for the design of diagnostic kits and U. urealyticum vaccine.
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spelling pubmed-98676102023-01-30 Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology Darvishi, Faleh Ganji, Ali Khansarinejad, Behzad Sadoogh Abbasian, Shabnam Abtahi, Hamid Iran J Microbiol Original Article BACKGROUND AND OBJECTIVES: The detection of Ureaplasma urealyticum is usually done through culture. With the change of the smallest effective factor in culture, we face the lack of growth of these bacteria, which is one of the important reasons to find a suitable alternative for the diagnosis of this bacterium. UreD is a protected gene in this bacterium. The aim of this was to evaluate the ability of antigenic regions of UreD protein to bind to patients’ serum antibodies. MATERIALS AND METHODS: Antigenic regions of UreD protein were predicted using IEDB software with five different methods: Emini Surface Accessibility Prediction, Kolaskar and Tongaonkar Antigenicity, Chou and Fasman beta turn prediction, Karplus and Schulz flexibility scale, Ellipro–Epitope prediction based upon structural protrusion. Antigenic regions of UreD gene was clonned, expressed and purified. The antigenicity of this recombinant protein against the antibodies in the serum of people infected with U. urealyticum infections was checked in western blotting. RESULTS: The results showed that the antigenic regions of the UreD protein was producted and its antigenicity was demonstrated in western blotting. Moreover, all sera from patients infected with U. urealyticum reacted to the recombinant antigen. CONCLUSION: Specimens from people infected with U. urealyticum infection was positive in Western blotting suggesting that UreD protein has antigenic properties. Therefore, it can be used as a suitable candidate for the design of diagnostic kits and U. urealyticum vaccine. Tehran University of Medical Sciences 2022-12 /pmc/articles/PMC9867610/ /pubmed/36721453 http://dx.doi.org/10.18502/ijm.v14i6.11255 Text en Copyright © 2022 The Authors. Published by Tehran University of Medical Sciences https://creativecommons.org/licenses/by-nc/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International license (https://creativecommons.org/licenses/by-nc/4.0/). Non-commercial uses of the work are permitted, provided the original work is properly cited.
spellingShingle Original Article
Darvishi, Faleh
Ganji, Ali
Khansarinejad, Behzad
Sadoogh Abbasian, Shabnam
Abtahi, Hamid
Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology
title Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology
title_full Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology
title_fullStr Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology
title_full_unstemmed Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology
title_short Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology
title_sort cloning, expression and purification of antigenic fragments of the ureaplasma urealyticum ured protein and its value in serology
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9867610/
https://www.ncbi.nlm.nih.gov/pubmed/36721453
http://dx.doi.org/10.18502/ijm.v14i6.11255
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