Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients

Abstract Background Circumsporozoite protein (CSP) has a central immune domain that includes short regions of repeating amino acid sequences. This immunodynamic region is an epitope of B cells that can elicit an immune response in human and laboratory animals. The aim of the present study was to exp...

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Autores principales:	Mehdi Nateghpour, Soudabeh Etemadi, Afsaneh Motevalli Haghi, Hamid Eslami, Mehdi Mohebali, Leila Farivar
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Plasmodium vivax Recombinant circumsporozoite protein VK210 ELISA Iran Medicine R
Acceso en línea:	https://doaj.org/article/c4829955b1434b438ada41ee4c82c5ca
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spelling	oai:doaj.org-article:c4829955b1434b438ada41ee4c82c5ca2021-11-28T12:23:39ZSerological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients10.1186/s40001-021-00607-62047-783Xhttps://doaj.org/article/c4829955b1434b438ada41ee4c82c5ca2021-11-01T00:00:00Zhttps://doi.org/10.1186/s40001-021-00607-6https://doaj.org/toc/2047-783XAbstract Background Circumsporozoite protein (CSP) has a central immune domain that includes short regions of repeating amino acid sequences. This immunodynamic region is an epitope of B cells that can elicit an immune response in human and laboratory animals. The aim of the present study was to express the recombinant PvCSP-VK210 antigen and evaluate it for assaying antibodies obtained during human P. vivax infection by Western blotting and indirect ELISA (enzyme-linked immunosorbent assay). Method Genomic DNA of P. vivax was isolated from a blood sample of an Iranian person with vivax malaria, and by PCR, the fragment of the PvCSP-VK210 gene was amplified. The gene fragment was cut after gel purification by BamHI and HindIII enzymes and then cloned into pET28a expression vector. Finally, the recombinant pET28a was transformed into the E. coli BL21 (DE3) as the expression host. In order to produce His-tagged protein, the expression host was cultured in LB medium. The protein was purified by Ni–NTA columns and immobilized metal affinity chromatography, and after confirmation by Western blotting technique, was used as the antigen in the indirect ELISA test. Results The recombinant protein was expressed and purified as a 32-kDa protein. The sensitivity and specificity of the indirect ELISA test with the recombinant PvCSP-VK210 antigen were 61.42% and 97.14%, respectively, based on OD = 0.313. Between the results of the microscopic test and the indirect ELISA test with the recombinant PvCSP-VK210 antigen there was a Kappa coefficient of 0.586. The positive and negative predictive value and validity of the ELISA test with the recombinant PvCSP-VK210 antigen were 95.55%, 71.57%, 79.28%, respectively. Conclusion The sensitivity of the indirect ELISA method with the recombinant PvCSP-VK210 antigen was 61.42%, which is the first report from Iran.Mehdi NateghpourSoudabeh EtemadiAfsaneh Motevalli HaghiHamid EslamiMehdi MohebaliLeila FarivarBMCarticlePlasmodium vivaxRecombinant circumsporozoite proteinVK210ELISAIranMedicineRENEuropean Journal of Medical Research, Vol 26, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Plasmodium vivax Recombinant circumsporozoite protein VK210 ELISA Iran Medicine R
spellingShingle	Plasmodium vivax Recombinant circumsporozoite protein VK210 ELISA Iran Medicine R Mehdi Nateghpour Soudabeh Etemadi Afsaneh Motevalli Haghi Hamid Eslami Mehdi Mohebali Leila Farivar Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients
description	Abstract Background Circumsporozoite protein (CSP) has a central immune domain that includes short regions of repeating amino acid sequences. This immunodynamic region is an epitope of B cells that can elicit an immune response in human and laboratory animals. The aim of the present study was to express the recombinant PvCSP-VK210 antigen and evaluate it for assaying antibodies obtained during human P. vivax infection by Western blotting and indirect ELISA (enzyme-linked immunosorbent assay). Method Genomic DNA of P. vivax was isolated from a blood sample of an Iranian person with vivax malaria, and by PCR, the fragment of the PvCSP-VK210 gene was amplified. The gene fragment was cut after gel purification by BamHI and HindIII enzymes and then cloned into pET28a expression vector. Finally, the recombinant pET28a was transformed into the E. coli BL21 (DE3) as the expression host. In order to produce His-tagged protein, the expression host was cultured in LB medium. The protein was purified by Ni–NTA columns and immobilized metal affinity chromatography, and after confirmation by Western blotting technique, was used as the antigen in the indirect ELISA test. Results The recombinant protein was expressed and purified as a 32-kDa protein. The sensitivity and specificity of the indirect ELISA test with the recombinant PvCSP-VK210 antigen were 61.42% and 97.14%, respectively, based on OD = 0.313. Between the results of the microscopic test and the indirect ELISA test with the recombinant PvCSP-VK210 antigen there was a Kappa coefficient of 0.586. The positive and negative predictive value and validity of the ELISA test with the recombinant PvCSP-VK210 antigen were 95.55%, 71.57%, 79.28%, respectively. Conclusion The sensitivity of the indirect ELISA method with the recombinant PvCSP-VK210 antigen was 61.42%, which is the first report from Iran.
format	article
author	Mehdi Nateghpour Soudabeh Etemadi Afsaneh Motevalli Haghi Hamid Eslami Mehdi Mohebali Leila Farivar
author_facet	Mehdi Nateghpour Soudabeh Etemadi Afsaneh Motevalli Haghi Hamid Eslami Mehdi Mohebali Leila Farivar
author_sort	Mehdi Nateghpour
title	Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients
title_short	Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients
title_full	Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients
title_fullStr	Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients
title_full_unstemmed	Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients
title_sort	serological responses to a soluble recombinant circumsporozoite protein-vk210 of plasmodium vivax (rpvcsp-vk210) among iranian malaria patients
publisher	BMC
publishDate	2021
url	https://doaj.org/article/c4829955b1434b438ada41ee4c82c5ca
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Serological responses to a soluble recombinant circumsporozoite protein-VK210 of Plasmodium vivax (rPvCSP-VK210) among Iranian malaria patients

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