Universidad Peruana Cayetano Heredia

A novel enolase from Taenia solium metacestodes and its evaluation as an immunodiagnostic antigen for porcine cysticercosis

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dc.contributor.author Ponce, Reynaldo
dc.contributor.author León-Janampa, Nancy
dc.contributor.author Gilman, Robert Hugh
dc.contributor.author Liendo, Ruddy
dc.contributor.author Roncal, Elisa
dc.contributor.author Luis, Sueline
dc.contributor.author Quiñones-Garcia, Stefany
dc.contributor.author Silverstein, Zach
dc.contributor.author García Lescano, Héctor Hugo
dc.contributor.author Gonzales, Armando
dc.contributor.author Sheen Cortavarria, Patricia
dc.contributor.author Zimic-Peralta, Mirko Juan
dc.contributor.author Pajuelo Travezaño, Monica Jhenny
dc.contributor.author Cysticercosis Working Group in Peru
dc.date.accessioned 2018-11-30T02:09:28Z
dc.date.available 2018-11-30T02:09:28Z
dc.date.issued 2018
dc.identifier.uri https://hdl.handle.net/20.500.12866/3985
dc.description.abstract Cysticercosis is a worldwide parasitic disease of humans and pigs principally caused by infection with the larvae of the pork tapeworm Taenia solium. Through the use of the recently-made-available T. solium genome, we identified a gene within a novel 1448 bp ORF that theoretically encodes for a 433 amino acid-long protein and predicted to be an α-enolase closely related to enolases of other flatworms. Additional bioinformatic analyses revealed a putative plasminogen-binding region on this protein, suggesting a potential role for this protein in pathogenesis. On this basis, we isolated the mRNA encoding for this presumptive enolase from T. solium metacestodes and reverse-transcribed it into cDNA before subsequently cloning and expressing it in both E. coli (rEnoTs) and insect cells (rEnoTsBac), in a 6xHis tagged manner. The molecular weights of these two recombinant proteins were ∼48 and ∼50 kDa, respectively, with the differences likely attributable to differential glycosylation. We used spectrophotometric assays to confirm the enolase nature of rEnoTs as well as to measure its enzymatic activity. The resulting estimates of specific activity (60.000 U/mg) and Km (0.091 mM) are quite similar to the catalytic characteristics of enolases of other flatworms. rEnoTs also exhibited high immunogenicity, eliciting a strong polyclonal antibody response in immunized rabbits. We subsequently employed rEnoTsBac for use in an ELISA aimed at discriminating between healthy pigs and those infected with T. solium. This diagnostic assay exhibited a sensitivity of 88.4% (95% CI, 74.92%-96.11%) and a specificity of 83.7% (95% CI: 69.29%-93.19%). In conclusión, this study reports on and enzymatically characterizes a novel enolase from T. solium metacestode, and shows a potential use as an immunodiagnostic for porcine cysticercosis. en_US
dc.language.iso eng
dc.publisher Elsevier
dc.relation.ispartofseries Experimental Parasitology
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Peru en_US
dc.subject Female en_US
dc.subject Cysticercosis en_US
dc.subject Taenia solium en_US
dc.subject Animals en_US
dc.subject Amino Acid Sequence en_US
dc.subject Antibodies, Helminth en_US
dc.subject Antigens, Helminth en_US
dc.subject Baculovirus en_US
dc.subject Computational Biology en_US
dc.subject Confidence Intervals en_US
dc.subject DNA, Complementary en_US
dc.subject E. coli en_US
dc.subject Enolase en_US
dc.subject Enzyme-Linked Immunosorbent Assay en_US
dc.subject Genetic Vectors en_US
dc.subject Immunodiagnosis en_US
dc.subject Phosphopyruvate Hydratase en_US
dc.subject Phylogeny en_US
dc.subject Protein Structure, Secondary en_US
dc.subject Protein Structure, Tertiary en_US
dc.subject Rabbits en_US
dc.subject Recombinant Proteins en_US
dc.subject RNA, Messenger en_US
dc.subject ROC Curve en_US
dc.subject Sensitivity and Specificity en_US
dc.subject Sequence Alignment en_US
dc.subject Sf9 Cells en_US
dc.subject Spectrophotometry en_US
dc.subject Swine en_US
dc.subject Swine Diseases en_US
dc.title A novel enolase from Taenia solium metacestodes and its evaluation as an immunodiagnostic antigen for porcine cysticercosis en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1016/j.exppara.2018.06.001
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.07
dc.relation.issn 1090-2449


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