title: Evaluation of baculovirus-derived recombinant 53-kDa protein of Trichinella spiralis for detection of Trichinella-specific antibodies in domestic pigs by ELISA.
abstract: The complete gene encoding the 53-kDa protein derived from Trichinella spiralis was cloned and expressed using a baculovirus-based system. Characterization of a purified fusion protein consisting of the 53-kDa protein and the glutathione S-transferase protein showed unspecific reactivity with swine pre-immune serum in both enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. Subsequently, a purified C-terminal 6xHis-tagged 53-kDa protein was used in an ELISA. The evaluation of the test using a negative serum panel showed a high specificity for the ELISA. Serum panels of pigs infected with T. spiralis of two independent experiments showed that pigs of one experiment were tested positive by the ELISA, whereas all sera of the second experiment were negative, indicating a low sensitivity of the ELISA. Furthermore, experimental evidence was found by using mass spectroscopy and Western blot analysis that the 53-kDa protein was not part of the excretory/secretory antigen of T. spiralis as shown in this study.
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
authors.forename: Doreen
authors.initials: D
authors.lastname: Jung
authors.forename: Jens Peter
authors.initials: JP
authors.lastname: Teifke
authors.forename: Axel
authors.initials: A
authors.lastname: Karger
authors.forename: Kathrin
authors.initials: K
authors.lastname: Michael
authors.forename: Simone
authors.initials: S
authors.lastname: Venz
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Wittmann
authors.forename: Katharina
authors.initials: K
authors.lastname: Kindermann
authors.forename: Karsten
authors.initials: K
authors.lastname: Nöckler
authors.forename: Egbert
authors.initials: E
authors.lastname: Mundt
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology / Q000302:isolation & purification*; D000947:Antigens, Helminth; D016367:Baculoviridae; D015415:Biomarkers; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D015801:Helminth Proteins; D007313:Insecta; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity; D013552:Swine / Q000276:immunology* / Q000469:parasitology*; D013553:Swine Diseases / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology; D017160:Trichinella spiralis / Q000276:immunology*; D014235:Trichinellosis / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015415:Biomarkers; D015801:Helminth Proteins; D011994:Recombinant Proteins