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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 H.
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, Hector H.
dc.contributor.author Gonzales, Armando
dc.contributor.author Sheen, Patricia
dc.contributor.author Zimic, Mirko
dc.contributor.author Pajuelo, Mónica J.
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 http://doi.org/10.1016/j.exppara.2018.06.001
dc.identifier.uri http://repositorio.upch.edu.pe/handle/upch/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.
dc.language.iso eng
dc.publisher Elsevier
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Peru
dc.subject Female
dc.subject Cysticercosis
dc.subject Taenia solium
dc.subject Animals
dc.subject Amino Acid Sequence
dc.subject Antibodies, Helminth
dc.subject Antigens, Helminth
dc.subject Baculovirus
dc.subject Computational Biology
dc.subject Confidence Intervals
dc.subject DNA, Complementary
dc.subject E. coli
dc.subject Enolase
dc.subject Enzyme-Linked Immunosorbent Assay
dc.subject Genetic Vectors
dc.subject Immunodiagnosis
dc.subject Phosphopyruvate Hydratase
dc.subject Phylogeny
dc.subject Protein Structure, Secondary
dc.subject Protein Structure, Tertiary
dc.subject Rabbits
dc.subject Recombinant Proteins
dc.subject RNA, Messenger
dc.subject ROC Curve
dc.subject Sensitivity and Specificity
dc.subject Sequence Alignment
dc.subject Sf9 Cells
dc.subject Spectrophotometry
dc.subject Swine
dc.subject Swine Diseases
dc.title A novel enolase from Taenia solium metacestodes and its evaluation as an immunodiagnostic antigen for porcine cysticercosis
dc.type info:eu-repo/semantics/article
dc.identifier.journal Experimental Parasitology

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