Type-specific serologic diagnosis of respiratory syncytial virus infection, based on a synthetic peptide of the attachment protein G.
Peptides deduced from the central hydrophobic region (residues 158-189) of the G protein of bovine and ovine respiratory syncytial virus (RSV) and of human RSV subtypes A and B were synthesized. These peptides were used to develop ELISAs to measure specifically antibodies against these types and subtypes of RSV. We have evaluated the bovine RSV-G peptide in both an indirect ELISA and in a blocking ELISA. Specificity and sensitivity, relative to a routine diagnostic ELISA that detects antibodies against the RSV F-protein in bovine sera, were 98% and 92% respectively for the indirect peptide-based ELISA, and 98% and 98% for the blocking peptide-based ELISA. In paired serum samples, rises in antibody titer were detected more frequently with the indirect peptide-based ELISA than with the routine F-ELISA. Furthermore, the peptide-based G-ELISAs were able to differentiate between antibodies against BRSV and HRSV, and between those against BRSV and ORSV. In addition, the indirect peptide-based ELISA was selective for HRSV subtype A and B antibodies. This study shows that peptides, corresponding to the central hydrophobic region of the attachment protein G of several RSVs, can be used successfully as antigens in highly specific and sensitive immunoassays.
Langedijk
J P
JP
Department of Bovine Virology, Institute for Animal Science and Health (ID-DLO), Lelystad, Netherlands. j.p.m.langedijk@id.dlo.nl
Middel
W G
WG
Schaaper
W M
WM
Meloen
R H
RH
Kramps
J A
JA
Brandenburg
A H
AH
van Oirschot
J T
JT
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D015707:HN Protein; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011817:Rabbits; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000821:virology; D012136:Respiratory Syncytial Viruses / Q000737:chemistry* / Q000145:classification*; D012703:Serotyping; D012756:Sheep; D013045:Species Specificity; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology*
D000956:Antigens, Viral; D015707:HN Protein; D010455:Peptides; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C116204:attachment protein G