Rapid methods for identification of poliovirus isolates and determination of polio neutralizing antibody titers in human sera.
A new rapid method for identification and determination of the titer of polioviruses and other enteroviruses in cell monolayers grown in microtiter plates is described. The method is based on immunoperoxidase staining of infected cells with commercial monoclonal antibodies (MAb) and biotin-labeled secondary antibody. The presence of poliovirus or other enteroviruses was established as the appearance of at least one focus of cells with stained cytoplasm 6 h post-infection. Viral titers determined by this method were expressed as focus forming unit (FFU) per ml which was found to correspond approximately to 10 TCID(50)/ml. The suitability of this technique to determine poliovirus antibody titers in human sera was also tested comparing the immunocytochemical neutralization assay with a conventional neutralization in microtiter plates. The test was standardized using reference human sera in order to produce antibody titers expressed in international units (IU). In addition to high reproducibility, the new neutralization test appears to be sensitive, specific and rapid, and might thus represent a useful tool for the diagnosis of polio and other enterovirus infections.
Di Lonardo
Anna
A
Laboratory of Virology, Istituto Superiore di Sanita', Viale Regina Elena 299, 00161, Rome, Italy.
Buttinelli
Gabriele
G
Amato
Concetta
C
Novello
Francesco
F
Ridolfi
Barbara
B
Fiore
Lucia
L
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood*; D019921:Biotinylation; D002460:Cell Line; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D009500:Neutralization Tests / Q000379:methods*; D011051:Poliomyelitis / Q000175:diagnosis / Q000821:virology*; D017955:Poliovirus / Q000276:immunology* / Q000302:isolation & purification*; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012703:Serotyping; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral