Time-resolved fluoroimmunoassays with monoclonal antibodies for rapid identification of parainfluenza type 4 and mumps viruses.
Monoclonal antibodies were prepared to the F and M proteins of parainfluenza 4A and 4B and to mumpsvirus to obtain reagents that could be configured into type-specific yet broadly-reactive IFA, EIA, and TR-FIA tests. Several antibodies to parainfluenza 4A also detected subtype 4B, although to a somewhat lower signal, and thus were well suited to generic parainfluenza type 4 tests that were comparable to similar tests previously described for parainfluenza types 1, 2, and 3. Monoclonals to subtype 4B were less able to detect 4A because of high background problems in one or another test. Monoclonals to mumpsvirus F protein were completely type-specific. These antibodies were screened by IFA and EIA for broad reactivity with diverse strains of each virus and were configured into optimized EIA and TR-FIA tests. The all-monoclonal tests were then compared to polyclonal tests in terms of their ability to detect virus in clinical specimens. The all-monoclonal TR-FIA was uniformly the most sensitive, detecting virus in 80% of culture-positive parainfluenza 4A specimens, 67% of parainfluenza 4B specimens, and 90% of mumps specimens, compared to 40-67% for the monoclonal EIA tests and 33-60% for the polyclonal EIA tests. For parainfluenza 4 TR-FIA, mean P/N values were 379 for subtype 4A cell culture fluids (228 for subtype 4B cultures) and 57 for 4A clinical specimens (43 for 4B specimens). For mumpsvirus TR-FIA, mean P/N values were 27 for culture fluids and 32 for clinical specimens. The sensitivities of the TR-FIA were determined with purified virus to be 0.28 ng virus per well for parainfluenza 4A and 0.70 ng virus per well for mumpsvirus.
Hierholzer
J C
JC
Division of Viral and Rickettsial Diseases, Center for Infectious Diseases, Centers for Disease Control, Atlanta, Georgia.
Bingham
P G
PG
Castells
E
E
Coombs
R A
RA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis; D015153:Blotting, Western; D003429:Cross Reactions; D005455:Fluorescent Antibody Technique; D015200:Fluoroimmunoassay / Q000379:methods*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009109:Mumps virus / Q000276:immunology / Q000302:isolation & purification*; D009666:Nose / Q000382:microbiology; D010614:Pharynx / Q000382:microbiology; D018108:Respirovirus / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral