Monoclonal Antibody Kit for Identification of the Novel 2009 H1N1 Influenza A Virus

Subtyping Influenza A Virus with Monoclonal Antibodies and an Indirect Immunofluorescence Assay

“…Finally, the 2009 H1 ID reagent stained the two 2009 H1N1 strains specifically with no cross-reactivity to 19 non-2009 H1N1 and H3N2 influenza A viruses ( Table 2). The subtyping reagents were further evaluated for use in shell vials on rhMK cells with selected 2009 H1N1, H1N1, and H3N2 influenza A virus strains following the standard IFA protocol as described previously (6). Results of this test were in 100% agreement with the previous tests on R-Mix monolayers (Fig.…”

“…Standard protocols for IFA and DFA were followed as described previously for detecting influenza A viruses and other prototype virus strains (6). To perform IFA against HRV, 96-well plates seeded with MRC-5 cells were infected with the virus strains.…”

“…A number of reports have described the development of subtype-specific MAbs, but they have not resulted in reagents for routine use (11,13,14,18). Recently we developed MAbs that exhibited specificity for the 2009 H1N1 pandemic influenza A virus, which formed the basis for a commercially available reagent for the identification of this virus (6). To extend this approach to other clinically relevant subtypes, we generated a series of subtype-specific MAbs for H3N2 and seasonal H1N1 subtypes as well and blended multiple subtype-specific MAbs to create indirect immunofluorescence assay (IFA) subtyping reagents.…”

Molecular Basis for Broad Neuraminidase Immunity: Conserved Epitopes in Seasonal and Pandemic H1N1 as Well as H5N1 Influenza Viruses

“…An immunofluorescence assay (IFA) was performed as described previously (28). Madin-Darby canine kidney (MDCK) cells growing in 96-well plates were infected with each virus at a multiplicity of infection of 1, followed by incubation at 35°C in 5% CO 2 for 48 h. Cells were fixed with acetone and ethanol (3:2, vol/vol) and incubated with each MAb, followed by incubation with fluorescein isothiocyanate (FITC)-conjugated anti-mouse IgG (Sigma-Aldrich, St. Louis, MO).…”

Diagnostic Methods of Influenza a From Clinical Specimens: A Critical Review of Literature

“…The first one is a mixture of A549 and Mink lung cells and the latter is a mixture of A549 and MDCK cell lines. During the 2009 H1N1 epidemics, the kit named D3 Ultra 2009 H1N1 Influenza A ID kit (2009 H1N1 ID kit; Diagnostic Hybrids, Inc., Ath¬ens, OH, USA) was developed using the R-mix Too cells allied to immunofluorescence method for the specific diagnosis of H1N1 ( Higgins et al 2010).…”