[The characterization of influenza viruses by the immunoperoxidase method using monoclonal antibodies. Its setup and validation].
The immunoperoxidase method for the rapid classification of influenza viruses in type and subtype was applied and validated for the first time in Cuba. The method is based on a rapid culture in MDCK-L cells and on the use of monoclonal antibodies for the classification in type and subtype. A pool of antibodies against influenza A and another against influenza B and HA1-71 and HA2-76 monoclonal antibodies are used for the subtyping in H1 and H3. The validation was carried out by applying this method to 21 international reference strains and to 23 human influenza virus strains that were isolated and previously classified by hemagglutination inhibition. All the strains reacted to the monoclonal antibodies according to their hemagglutinin type and subtype. 6 reference strains and 9 isolations were characterized within the H1N1 subtype: 9 reference strains and 10 isolations in the H3N2 subtype; and 6 reference strains and 4 isolations in type B. There were neither unspecific nor crossed reactions among the controls established. There was 100% of sensitivity, specificity and coincidence. The technique used proved to be fast and convenient for the characterization in type and subtype of the isolated influenza virus strains. It may substitute the classic hemagglutination inhibition method when it is required the rapid characterization of outbreaks or epidemics of acute respiratory infections, which is very important due to the high morbidity they cause mainly in risk groups and to their economic repercussion.
Oropesa Fernández
S
S
Instituto de Medicina Tropical Pedro Kourí.
Morier Díaz
L
L
Vázquez Ramudo
S
S
Goyenechea Hernández
A
A
Valdivia Romero
A
A
Hernández Espinosa
B
B
García Infante
S
S
D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D002478:Cells, Cultured; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods / Q000592:standards; D009980:Influenza A virus / Q000145:classification* / Q000276:immunology; D009981:Influenza B virus / Q000145:classification* / Q000276:immunology; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods / Q000592:standards; D014776:Virus Cultivation
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral