Evaluation of baculovirus-derived recombinant 53-kDa protein of Trichinella spiralis for detection of Trichinella-specific antibodies in domestic pigs by ELISA

Jung, Doreen; Teifke, Jens Peter; Karger, Axel; Michael, Kathrin; Venz, S.; Wittmann, W.; Kindermann, K.; Nöckler, K.; Mundt, Egbert

doi:10.1007/s00436-006-0296-7

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Evaluation of baculovirus-derived recombinant 53-kDa protein of Trichinella spiralis for detection of Trichinella-specific antibodies in domestic pigs by ELISA

Jung, Doreen ; Teifke, Jens Peter ; Karger, Axel ; Michael, Kathrin ; Venz, S.; Wittmann, W.; Kindermann, K.; Nöckler, K.; Mundt, Egbert

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

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Erschienen in:: Parasitology Research
Vol. 100, H. 3 (2007), S. 429-437
Band:: 100
Heft:: 3
Datum der Veröffentlichung:: 2007
DOI:: 10.1007/s00436-006-0296-7
Sprache:: Englisch
Ressourcentyp:: Text
Schlagwörter:: analysis; animal; Animals; Antibodies; Antibodies,Helminth; ANTIBODY; ANTIGEN; ANTIGENS; Antigens,Helminth; article; ASSAY; Baculoviridae; Biological Markers; Cell Line; detection; diagnosis; ELISA; Enzyme-Linked Immunosorbent Assay; evaluation; Evaluation Studies; gene; genetics; Germany; GLUTATHIONE-S-TRANSFERASE; HEALTH; Helminth Proteins; immunology; Insects; isolation & purification; MARKERS; parasitology; pig; PIGS; protein; Proteins; recombinant proteins; Research; SENSITIVITY; Sensitivity and Specificity; SERA; serum; SPECIFICITY; SPIRALIS; studies; study; Swine; Swine Diseases; SYSTEM; Trichinella; Trichinella spiralis; TRICHINOSIS; veterinary; western blot; WESTERN-BLOT-ANALYSIS
DDC-Sachgruppe der DNB:: 600 Technik
Link URL:: http://link.springer.com/article/10.1007%2Fs00436-006-0296-7
Einrichtung:: Bundesinstitut für Risikobewertung, Bundesinstitut für Risikobewertung (Juli 2014 - 2021), Abteilung 4 - Biologische Sicherheit (Juli 2014 - 2021), Fachgruppe 45 - Diagnostik und Erregercharakterisierung (Juli 2014 - 2021)
Einrichtung:: Friedrich-Loeffler-Institut, Institut für molekulare Virologie und Zellbiologie
Einrichtung:: Friedrich-Loeffler-Institut, Institut für Infektionsmedizin
Physischer Standort:: SD/2007/150