Influenza Virus Typing Set
Applications:
- Typing of Influenza Virus A and B Subtypes
Description:
Influenza viruses are classified into three groups (A, B and C) based upon differences for their nuclear proteins (NP), and are further classified into subtypes according to their surface glycoproteins, neuraminidase (NA) and haemagglutinin (HA). The most prevalent types of influenza virus are influenza A H1, influenza A H3, and influenza B. The influenza A HA protein consists of a head region which contains a target cell receptor domain, and a stem region that contains a special amino acid sequence required for fusion of the virus and the target cell. The HA head region, in particular, has especially strong antigenicity and, thus, most produced antibodies recognize this region. However, mutation readily occurs in this region, and typing represents a significant method for judging the effect of a vaccination. Furthermore, rapid typing is important due to the short infectious period of this virus.
The Influenza Virus Typing Set allows typing of influenza virus A and B subtypes. The Set contains several biotinylated antibodies (two kinds of monoclonal antibodies that distinguish between influenza A subtypes, one monoclonal antibody that binds all types of influenza A viruses, one polyclonal antibody that binds all types of A and B viruses) and Avidin-POD. The sensitivity of this system is equivalent to that of the PAP method.
The protocol involves preparing a virus solution from samples, which is then used to infect Madin-Darby Canine Kidney Epithelial Cells (MDCK) cells in 96-well plates. The culture supernatant is removed, cells are ethanol-fixed, and an avidin solution is added to each well. Following blocking of non-specific binding regions, a biotinylated antibody is added to the well, followed by addition of the streptavidin POD conjugate. Finally, the well is washed and the coloring substrate Diaminobenzidine (DAB, not supplied) is added. Positive subtype determination is made based upon color development within the well.
Kit Components:
| Biotinylated-C179 Antibody | 1 x 100 μL |
| Biotinylated-F49 Antibody | 1 x 100 μL |
| Biotinylated-C111 Antibody | 1 x 100 μL |
| Biotinylated-9D6 Antibody | 1 x 100 μL |
| Biotinylated-Anti Influenza A, B | 1 x 100 μL |
| Avidin-POD | 1 x 500 μL |
| Diluent (for antibody, X 4) | 1 x 250 ml |
Antibodies Supplied with this Kit
Biotinylated-C179: Human Influenza A Monoclonal Antibody (H1N1,H2N2) (Cat.# TAK M145). This antibody specifically recognizes the conformational epitope in the stem region of HA in which amino acid sequences are highly conserved in H1N1 and H2N2 subtypes of Influenza A virus.
Biotinylated-F49: Human Influenza A Monoclonal antibody (H3N2) (Cat.# TAK M146). This antibody specifically recognizes the stem region of HA in H3N2 subtype of Influenza A virus.
Biotinylated-C111: Human Influenza A Monoclonal antibody (H1, H2, H3) (Cat.# TAK M147). This antibody specifically recognizes the matrix protein in H1N1, H2N2 and H3N2 all subtypes of Influenza A virus.
Biotinylated-9D6: Human Influenza B Monoclonal antibody to Cat.# TAK M148). This antibody specifically recognizes the influenza B virus nuclear protein (NP).
Biotinylated-anti influenza A, B: Polyclonal antibody to human influenza A, B (Cat.# TAK M149). This antibody was created by using a vaccine mixture of Influenza A/Peking/262/95 (H1N1), A/Wuhan/359/95 (H3N2), B/Mie/1/93 and B/Guandong/5/94 for an immunogen. This antibody reacts equally with both Influenza A and B virus.
Performance Characteristics
| Assay sensitivity: | Equivalent to PAP method |
| Assay duration: | 1.5 hours |
| Specificity: | Influenza virus A and B subtypes |
| Test specimen type: | MDCK cells infected with the sample virus |
