Antigen-capture blocking enzyme-linked immunosorbent assay based on a baculovirus recombinant antigen to differentiate Transmissible gastroenteritis virus from Porcine respiratory coronavirus antibodies.
A new commercially available antigen-capture, blocking enzyme-linked immunosorbent assay (antigen-capture b-ELISA), based on baculovirus truncated-S recombinant protein of Transmissible gastroenteritis virus (TGEV) and 3 specific monoclonal antibodies, was developed and evaluated by examining a panel of 453 positive Porcine respiratory coronavirus (PRCoV), 31 positive TGEV, and 126 negative field sera by using another commercially available differential coronavirus b-ELISA as the reference technique to differentiate TGEV- from PRCoV-induced antibodies. The recombinant S protein-based ELISA appeared to be 100% sensitive for TGEV and PRCoV detection and highly specific for TGEV and PRCoV detection (100% and 92.06%, respectively), when qualitative results (positive or negative) were compared with those of the reference technique. In variability experiments, the ELISA gave consistent results when the same serum was evaluated on different wells and different plates. These results indicated that truncated recombinant S protein is a suitable alternative to the complete virus as antigen in ELISA assays. The use of recombinant S protein as antigen offers great advantages because it is an easy-to-produce, easy-to-standardize, noninfectious antigen that does not require further purification or concentration. Those advantages represent an important improvement for antigen preparation, in comparison with other assays in which an inactivated virus from mammalian cell cultures is used.
López
Lissett
L
Inmunología y Genética Aplicada S.A. (INGENASA), Hnos. García Noblejas 41, 28037 Madrid, Spain.
Venteo
Angel
A
García
Marga
M
Camuñas
Ana
A
Ranz
Ana
A
García
Julia
J
Sarraseca
Javier
J
Anaya
Carmen
C
Rueda
Paloma
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D017925:Circovirus / Q000276:immunology; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D009500:Neutralization Tests; D028323:Parvovirus, Porcine / Q000276:immunology; D019318:Porcine Reproductive and Respiratory Syndrome / Q000276:immunology / Q000401:mortality; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000276:immunology*; D017293:Protein S / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018411:Spodoptera / Q000276:immunology; D013552:Swine
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D017293:Protein S