Quantification of respiratory syncytial virus polypeptides in nasal secretions by monoclonal antibodies.
An indirect enzyme-linked immunosorbent assay (ELISA) which uses monoclonal antibody as solid-phase immunosorbent was developed to measure specific polypeptides of respiratory syncytial virus (RSV). The assay was used to examine 43 nasopharyngeal (NP) aspirates from RSV-positive infants that had been examined previously for RSV by culture, direct immunofluorescence, and polyclonal antibody ELISA. Frozen NP aspirates were serially diluted and examined for the 66K mol. wt. fusion glycoprotein (F), the 84K large surface glycoprotein (G) and the 41K nucleoprotein (N) by monoclonal capture ELISA. F protein was detected in all 43 specimens, G protein was detectable in 20 (47%) and N protein in 22 (51%) of 43 NP aspirates. In specimens with detectable G and N proteins, F was detected by endpoint titration at approximately tenfold greater dilutions than either G or N. In 19 sequential NP aspirates from five patients with RSV infection, F was present in higher titre throughout infection. In 20 cases, matching cell culture isolates were examined by immunofluorescence with strain-specific monoclonal antibodies. Three of 20 isolates showed strain-specific differences by their lack of reaction with anti-G monoclonal antibody. Titration of the 20 cell culture isolates by monoclonal antibody capture ELISA showed the relative amount of F and N proteins to be equal in all cases, whereas levels of G protein tended to be slightly lower. Reconstruction experiments with NP aspirates demonstrated that degradation of F and N proteins did not occur in NP aspirates, but that G protein antigenicity appeared to be affected by nasal secretions. When compared with cell culture-grown material, nasal secretions contained abundant F protein but a surprisingly low concentration of N protein.
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D009297:Nasal Mucosa / Q000378:metabolism / Q000382:microbiology*; D010455:Peptides / Q000032:analysis*; D012136:Respiratory Syncytial Viruses / Q000302:isolation & purification*; D014764:Viral Proteins / Q000032:analysis*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D010455:Peptides; D014764:Viral Proteins