Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus

Abstract The larval stage of Echinococcus granulosus causes cystic echinococcosis (CE), a neglected tropical disease that leads to morbidity and mortality in humans and livestock worldwide. Here, we identified and characterized dihydrofolate reductase (Eg-DHFR) from E. granulosus, and evaluated its...

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Autores principales:	Xingju Song, Dandan Hu, Min Yan, Yu Wang, Ning Wang, Xiaobin Gu, Guangyou Yang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/35d1753521f4437ea149c88b286026bc
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spelling	oai:doaj.org-article:35d1753521f4437ea149c88b286026bc2021-12-02T12:30:25ZMolecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus10.1038/s41598-017-00643-52045-2322https://doaj.org/article/35d1753521f4437ea149c88b286026bc2017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00643-5https://doaj.org/toc/2045-2322Abstract The larval stage of Echinococcus granulosus causes cystic echinococcosis (CE), a neglected tropical disease that leads to morbidity and mortality in humans and livestock worldwide. Here, we identified and characterized dihydrofolate reductase (Eg-DHFR) from E. granulosus, and evaluated its potential as a diagnostic antigen for sheep CE. Comparison between mammalian (host) DHFR and Eg-DHFR indicates that 45.7% of the 35 active site residues are different. Immunolocalisation analysis showed that native Eg-DHFR was widely distributed in all life-cycle stages of E. granulosus. Recombinant Eg-DHFR (rEg-DHFR) showed typical DHFR enzymatic parameters towards substrate, and was very sensitive to inhibition by methotrexate (IC50 = 27.75 ± 1.03 nM) and aminopterin (IC50 = 63.67 ± 6.76 nM). However, inhibition of DHFR exhibited little protoscolicidal effect in vitro. As there is no reliable method to monitor sheep CE, the immunogenicity of rEg-DHFR was detected, and we developed an indirect ELISA (iELISA) for CE serodiagnosis. The iELISA exhibited diagnostic specificity of 89.58%, diagnostic sensitivity of 95.83%, and the diagnostic accuracy was 91.67% compared with necropsy. Cross-reactivity assay showed analytical specificity of 85.7%. These suggest that rEg-DHFR is an effective antigen for the diagnosis of sheep CE.Xingju SongDandan HuMin YanYu WangNing WangXiaobin GuGuangyou YangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Xingju Song Dandan Hu Min Yan Yu Wang Ning Wang Xiaobin Gu Guangyou Yang Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus
description	Abstract The larval stage of Echinococcus granulosus causes cystic echinococcosis (CE), a neglected tropical disease that leads to morbidity and mortality in humans and livestock worldwide. Here, we identified and characterized dihydrofolate reductase (Eg-DHFR) from E. granulosus, and evaluated its potential as a diagnostic antigen for sheep CE. Comparison between mammalian (host) DHFR and Eg-DHFR indicates that 45.7% of the 35 active site residues are different. Immunolocalisation analysis showed that native Eg-DHFR was widely distributed in all life-cycle stages of E. granulosus. Recombinant Eg-DHFR (rEg-DHFR) showed typical DHFR enzymatic parameters towards substrate, and was very sensitive to inhibition by methotrexate (IC50 = 27.75 ± 1.03 nM) and aminopterin (IC50 = 63.67 ± 6.76 nM). However, inhibition of DHFR exhibited little protoscolicidal effect in vitro. As there is no reliable method to monitor sheep CE, the immunogenicity of rEg-DHFR was detected, and we developed an indirect ELISA (iELISA) for CE serodiagnosis. The iELISA exhibited diagnostic specificity of 89.58%, diagnostic sensitivity of 95.83%, and the diagnostic accuracy was 91.67% compared with necropsy. Cross-reactivity assay showed analytical specificity of 85.7%. These suggest that rEg-DHFR is an effective antigen for the diagnosis of sheep CE.
format	article
author	Xingju Song Dandan Hu Min Yan Yu Wang Ning Wang Xiaobin Gu Guangyou Yang
author_facet	Xingju Song Dandan Hu Min Yan Yu Wang Ning Wang Xiaobin Gu Guangyou Yang
author_sort	Xingju Song
title	Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus
title_short	Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus
title_full	Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus
title_fullStr	Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus
title_full_unstemmed	Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus
title_sort	molecular characteristics and serodiagnostic potential of dihydrofolate reductase from echinococcus granulosus
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/35d1753521f4437ea149c88b286026bc
work_keys_str_mv	AT xingjusong molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus AT dandanhu molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus AT minyan molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus AT yuwang molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus AT ningwang molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus AT xiaobingu molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus AT guangyouyang molecularcharacteristicsandserodiagnosticpotentialofdihydrofolatereductasefromechinococcusgranulosus
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Molecular Characteristics and Serodiagnostic Potential of Dihydrofolate Reductase from Echinococcus granulosus

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