Trypanosoma cruzi flagellar repetitive antigen expression by recombinant baculovirus: towards an improved diagnostic reagent for Chagas' disease

Biotechnology (N Y). 1992 Nov;10(11):1474-7. doi: 10.1038/nbt1192-1474.

Abstract

We constructed a recombinant baculovirus that expressed part of a Trypanosoma cruzi flagellar repetitive antigen (FRA). Both cell- associated and secreted forms of recombinant FRA were detected in cultures of virus-infected Spodoptera frugiperda (Sf9) cells. These forms show a complex pattern after polyacrylamide gel electrophoresis and Western blot analysis using either an anti-FRA rabbit serum or human Chagasic sera. Competitive Western-blot experiments revealed that all bands react with the same antibodies as a bacterially-derived FRA. Polymerase chain reaction and Southern blots of the recombinant viral DNA also showed a complex pattern, suggesting the presence of more than one repeat unit in the viral genome. When tested against a panel of human sera from an endemic area for Chagas' disease, FRA recombinant-Sf9 culture supernatant showed the same reactivity as purified FRA produced in bacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Protozoan / immunology*
  • Baculoviridae / immunology*
  • Biotechnology*
  • Cells, Cultured
  • Chagas Disease / diagnosis*
  • DNA, Viral / analysis
  • Enzyme-Linked Immunosorbent Assay
  • Flagella / immunology*
  • Immunoassay
  • Insecta / cytology
  • Polymerase Chain Reaction
  • Trypanosoma cruzi
  • Viral Proteins / analysis

Substances

  • Antigens, Protozoan
  • DNA, Viral
  • Viral Proteins