tetano
Editor, Senior Moderator
J Clin Microbiol. 2010 Jun 2. [Epub ahead of print]
A Monoclonal Antibody Kit for Identification of the Novel 2009 H1N1 Influenza A Virus.
Higgins AD, Shaw CJ, Johnson JG, Navarro A, Chapman NA, Ewers SD, Stockwell JW, Carpenter JM, Olivo PD, Miao LY.
Diagnostic Hybrids, Inc., 1055 East State St., Athens, Ohio 45701.
Abstract
To develop an immunofluorescence assay for identification of the 2009 H1N1 influenza A virus, we generated a number of monoclonal antibodies (MAbs) using inactivated H1N1 2009 virus (A/California/07/2009) as the immunogen. Two MAbs that target two different epitopes of the 2009 H1N1 hemagglutinin were selected to make the D3 Ultra 2009 H1N1 Influenza A ID Kit (2009 H1N1 ID Kit, Diagnostic Hybrids, Inc., Athens, OH) that is intended for the identification of the 2009 H1N1 virus by indirect immunofluorescence assay (IFA). The kit does not detect any of 14 seasonal H1N1 or H3N2 influenza prototype virus strains, and is also not reactive with seven other major respiratory viruses. Clinical respiratory specimens were evaluated using both the 2009 H1N1 ID Kit and the CDC Human Influenza Virus Real-time RT-PCR Swine Flu Panel (CDC rRT-PCR) and showed 100% agreement between the two assays. Four of these clinical specimens, however, were positive by the 2009 H1N1 ID Kit, but were identified as presumptively positive by the CDC rRT-PCR by virtue of showing Ct values only with universal InfA and swInfA primers, but not with swH1 primers. Sequence analysis of the HA gene in these four specimens revealed point mutations in both primer and probe regions. In addition, unlike the CDC rRT-PCR, the 2009 H1N1 ID Kit can differentiate the 2009 H1N1 virus from a swine-derived H1 influenza A virus (A/New Jersey/8/76). The 2009 H1N1 ID Kit offers clinical laboratories an alternative to RT-PCR for the identification of 2009 H1N1 influenza A virus.
PMID: 20519459 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/20519459
A Monoclonal Antibody Kit for Identification of the Novel 2009 H1N1 Influenza A Virus.
Higgins AD, Shaw CJ, Johnson JG, Navarro A, Chapman NA, Ewers SD, Stockwell JW, Carpenter JM, Olivo PD, Miao LY.
Diagnostic Hybrids, Inc., 1055 East State St., Athens, Ohio 45701.
Abstract
To develop an immunofluorescence assay for identification of the 2009 H1N1 influenza A virus, we generated a number of monoclonal antibodies (MAbs) using inactivated H1N1 2009 virus (A/California/07/2009) as the immunogen. Two MAbs that target two different epitopes of the 2009 H1N1 hemagglutinin were selected to make the D3 Ultra 2009 H1N1 Influenza A ID Kit (2009 H1N1 ID Kit, Diagnostic Hybrids, Inc., Athens, OH) that is intended for the identification of the 2009 H1N1 virus by indirect immunofluorescence assay (IFA). The kit does not detect any of 14 seasonal H1N1 or H3N2 influenza prototype virus strains, and is also not reactive with seven other major respiratory viruses. Clinical respiratory specimens were evaluated using both the 2009 H1N1 ID Kit and the CDC Human Influenza Virus Real-time RT-PCR Swine Flu Panel (CDC rRT-PCR) and showed 100% agreement between the two assays. Four of these clinical specimens, however, were positive by the 2009 H1N1 ID Kit, but were identified as presumptively positive by the CDC rRT-PCR by virtue of showing Ct values only with universal InfA and swInfA primers, but not with swH1 primers. Sequence analysis of the HA gene in these four specimens revealed point mutations in both primer and probe regions. In addition, unlike the CDC rRT-PCR, the 2009 H1N1 ID Kit can differentiate the 2009 H1N1 virus from a swine-derived H1 influenza A virus (A/New Jersey/8/76). The 2009 H1N1 ID Kit offers clinical laboratories an alternative to RT-PCR for the identification of 2009 H1N1 influenza A virus.
PMID: 20519459 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/20519459