Detection of antibodies against foot-and-mouth disease virus using a liquid-phase blocking sandwich ELISA (LPBE) with a bioengineered 3D protein.
A liquid-phase blocking sandwich enzyme-linked immunosorbent assay (ELISA-3D) was developed to detect specific antibodies to the 3D protein in sera from foot-and-mouth disease (FMD) virus (FMDV)-infected animals. The assay uses a nonstructural 3D recombinant protein and two polyclonal antisera, one for capture (bovine) and the other for detector (guinea pig). The specificity of the assay was demonstrated by negative results with 101 sera of cattle from the FMD-free zone in Argentina and with bovine and porcine sera raised against various RNA and DNA viruses. The ELISA-3D was able to detect antibodies in cattle after natural or experimental infection with FMDV of A, O, or C types as early as 5 days postinfection and at later stages in persistently infected animals. Comparison of the results with those obtained with the routinely used agar gel immunodiffusion test and a previously described ELISA, both employing a partially purified virus-infection-associated antigen, shows that the ELISA-3D is highly sensitive and specific and gives reproducible results. Its use as a tool for monitoring viral activity and for certification of FMDV-free animals is recommended.
O'Donnell
V K
VK
Instituto de Virologia, Centro de Investigación en Ciencias Veterinarias, INTA, Morón, Provincia de Buenos Aires, Argentina.
Boyle
D B
DB
Sproat
K
K
Fondevila
N A
NA
Forman
A
A
Schudel
A A
AA
Smitsaart
E N
EN
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D005537:Aphthovirus / Q000276:immunology; D001118:Argentina; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005536:Foot-and-Mouth Disease / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D005982:Glutathione Transferase; D006168:Guinea Pigs; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D013552:Swine; D013997:Time Factors; D014611:Vaccination; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014765:Viral Vaccines
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C103557:virus-infection-associated antigen, Foot-and-mouth disease virus; D005982:Glutathione Transferase