Truncated E2 of bovine viral diarrhea virus (BVDV) expressed in Drosophila melanogaster cells: a candidate antigen for a BVDV ELISA.
A simple and reliable indirect enzyme-linked immunosorbent assay for detection of antibodies directed against a major bovine viral diarrhea virus (BVDV) immunogen, the E2 glycoprotein (tE2-ELISA), has been developed using the recombinant C-terminal truncated E2 glycoprotein (tE2) expressed in a Drosophila melanogaster system. This strategy demonstrated that tE2 is secreted efficiently in the supernatant, no purification steps are necessary, it is easy to produce and carries out the post translational modifications necessary to preserve its native conformation. Preliminary analysis of 183 cattle serum samples using tE2-ELISA showed a 98% specificity and a 100% sensitivity compared with the standard homologous BVDV virus neutralization test. The results also showed that the tE2 is immunoreactive because the conformation and antigenicity of the original E2 are maintained to a large extent. To our knowledge this is the first study report of the recombinant tE2 of BVDV expressed in D. melanogaster system as an antigen for ELISA.
Marzocca
M P
MP
Fundación de estudios en Virología Animal (FEVAN), Guamini 1682, Ciudad de Buenos Aires (C1440ESD), Argentina.
Seki
C
C
Giambiagi
S M
SM
Robiolo
B
B
Schauer
R
R
Dus Santos
M J
MJ
Scodeller
E A
EA
La Torre
J L
JL
Wigdorovitz
A
A
Grigera
P R
PR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D002417:Cattle; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics / Q000276:immunology*; D004331:Drosophila melanogaster; D004797:Enzyme-Linked Immunosorbent Assay; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D014765:Viral Vaccines / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines