[Fast detection and characterization of influenza A and B viruses in nasopharyngeal secretions by the immunoperoxidase method]. / Detección y caracterización rápida de los virus de influenza A y B en secreciones nasofaríngeas mediante el método de la inmunoperoxidasa.

One hundred and fourty eight samples from patients with a symptomatology compatible with the influenza virus were studied aimed at identifying in a fast way these viruses. A rapid MDCK-L cell culture was developed on 96 well plates, where nasopharingeal exudates or gargarisms were inoculated and incubated all night long at 37 degrees C. The medium was removed and cells were washed with PBS and fixed with methanol. Viral antigens were detected through the immunoperoxidase staining by using two monoclonal antibody pools for the identification of influenza A and influenza B viruses. The HA1-71 monoclonal antibody, specific for influenza A (H3N2) and the HA2-76, which react with both A (H3N2) and A (H1N1) were used for subtyping. Of all the positive samples (136), 72% corresponded to type A while 34.6% and 37.5% corresponded to subtypes H1 and H3, respectively. Influenza B was detected in 27.9% of the 148 samples studied. Only 12 were negative (8.1%). The use of this technique is recommended as a rapid, convenient and sensitive method that is easy to carry put and to interpretate for the detection and characterization in type and subtype of the influenza viruses starting from the nasopharyngeal exudates or gargarisms.

[The characterization of influenza viruses by the immunoperoxidase method using monoclonal antibodies. Its setup and validation]. / Caracterización de los virus de influenza por el método de la inmunoperoxidasa utilizando anticuerpos monoclonales. Montaje y validación.

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.