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J Clin Microbiol. 2011 Nov 9. [Epub ahead of print]
Subtyping influenza A Virus with Monoclonal Antibodies and an Indirect Immunofluorescence Assay.
Johnson J, Higgins A, Navarro A, Huang Y, Esper FL, Barton N, Esch D, Shaw C, Olivo PD, Miao LY.
Source
Diagnostic Hybrids, Inc. 1055 East State St. Athens, Ohio 45701.
Abstract
The recent association of certain Influenza A subtypes with clinically relevant phenotypes has led to the increasing importance of subtyping by clinical virology laboratories. To provide clinical laboratories with a definitive immunofluorescence assay for the subtyping of influenza A virus isolates, we generated a panel of monoclonal antibodies (MAbs) against the major circulating influenza A subtypes using multiple inactivated H1N1, H3N2, and 2009 H1N1 strains individually as immunogens. Eleven MAbs that target hemagglutinin (HA) of H1N1 and H3N2 subtypes were selected. These MAbs were combined into three subtype-specific reagents, one each for pan-H1 (seasonal and 2009 strains), H3, and 2009 H1for the subtyping of influenza A-positive specimens by indirect immunofluorescence assay (IFA). Each subtype-specific reagent was tested on twenty-one prototype influenza A virus strains and confirmed to be specific for its intended subtype. In addition, the subtyping reagents did not cross-react with any of forty other viruses. The clinical performance of the subtyping reagents was evaluated with seventy-five archived clinical samples collected between 2006 and 2009 using the D(3) Ultra DFA influenza A ID Reagent (Diagnostic Hybrids, Inc., a Quidel Company, Athens, OH) and the influenza A subtyping reagents by IFA simultaneously. Sixty-four samples grew virus and were subtyped as follows: thirty H3N2, nine seasonal H1N1 and twenty-five 2009 H1N1. RT-PCR was used to confirm subtyping influenza A of these samples and there was 100% agreement with IFA. This subtyping IFA provides clinical laboratories with a cost-effective, diagnostic tool for better management of influenza virus infection and surveillance of influenza virus activity.
PMID:
22075584
[PubMed - as supplied by publisher]
Subtyping influenza A Virus with Monoclonal Antibodies and an Indirect Immunofluorescence Assay.
Johnson J, Higgins A, Navarro A, Huang Y, Esper FL, Barton N, Esch D, Shaw C, Olivo PD, Miao LY.
Source
Diagnostic Hybrids, Inc. 1055 East State St. Athens, Ohio 45701.
Abstract
The recent association of certain Influenza A subtypes with clinically relevant phenotypes has led to the increasing importance of subtyping by clinical virology laboratories. To provide clinical laboratories with a definitive immunofluorescence assay for the subtyping of influenza A virus isolates, we generated a panel of monoclonal antibodies (MAbs) against the major circulating influenza A subtypes using multiple inactivated H1N1, H3N2, and 2009 H1N1 strains individually as immunogens. Eleven MAbs that target hemagglutinin (HA) of H1N1 and H3N2 subtypes were selected. These MAbs were combined into three subtype-specific reagents, one each for pan-H1 (seasonal and 2009 strains), H3, and 2009 H1for the subtyping of influenza A-positive specimens by indirect immunofluorescence assay (IFA). Each subtype-specific reagent was tested on twenty-one prototype influenza A virus strains and confirmed to be specific for its intended subtype. In addition, the subtyping reagents did not cross-react with any of forty other viruses. The clinical performance of the subtyping reagents was evaluated with seventy-five archived clinical samples collected between 2006 and 2009 using the D(3) Ultra DFA influenza A ID Reagent (Diagnostic Hybrids, Inc., a Quidel Company, Athens, OH) and the influenza A subtyping reagents by IFA simultaneously. Sixty-four samples grew virus and were subtyped as follows: thirty H3N2, nine seasonal H1N1 and twenty-five 2009 H1N1. RT-PCR was used to confirm subtyping influenza A of these samples and there was 100% agreement with IFA. This subtyping IFA provides clinical laboratories with a cost-effective, diagnostic tool for better management of influenza virus infection and surveillance of influenza virus activity.
PMID:
22075584
[PubMed - as supplied by publisher]
