Development of an Immunochromatographic Assay Specifically Detecting Pandemic H1N1 (2009) Influenza Virus

Abstract

The pandemic caused by a new type of influenza virus, pandemic H1N1 (2009) influenza virus A (AH1pdm), has had a major worldwide impact. Since hemagglutinin (HA) genes are among the most specific genes in the influenza virus genome, AH1pdm can be definitively diagnosed by viral gene analysis targeting the HA genes. This type of analysis, however, cannot be easily performed in clinical settings. While commercially available rapid diagnosis kits (RDKs) based on immunochromatography can be used to detect nucleoproteins (NPs) of influenza A and B viruses in clinical samples, there are no such kits that are specific for AH1pdm. We show here that an RDK using a combination of monoclonal antibodies against NP can be used to specifically detect AH1pdm. The RDK recognized AH1pdm virus isolates but did not recognize seasonal H1N1 and H3N2 and influenza B viruses, indicating that the specificity of the RDK is 100%. A parallel comparison of RDK with a commercial influenza A/B virus kit revealed that both types of kits had equal sensitivities in detecting their respective viruses. Preliminary evaluation of clinical samples from 5 individuals with PCR-confirmed human AH1pdm infection showed that the RDK was positive for all samples, with the same detection intensity as that of a commercial influenza A/B virus kit. This RDK, together with a new vaccine and the stockpiling of anti-influenza drugs, will make aggressive measures to contain AH1pdm infections possible.

Development of Two Types of Rapid Diagnostic Test Kits To Detect the Hemagglutinin or Nucleoprotein of the Swine-Origin Pandemic Influenza A Virus H1N1

“…MiyoshiAkiyama et al (17) developed a test kit that can be used to distinguish H1N1 pdm from seasonal influenza A (H1N1 and H3N2) and B viruses. They used an anti-NP MAb obtained from mice immunized with H5N1 and evaluated it using five clinical specimens.…”

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

“…The 2009 pandemic further emphasized the need for clinical laboratories to be able to subtype viruses. Recently, a report was published on two MAbs that were generated using a recombinant nucleoprotein of H5N1 virus and serendipitously were found to react with the 2009 H1N1 virus, without apparent cross-reactivity with other human seasonal influenza A viruses (10). These MAbs were used in an immunochromatographic assay for specifically detecting and identifying the 2009 H1N1 virus.…”

Immunological characterization of monoclonal antibodies used in rapid influenza diagnostic test for detection of the 2009 pandemic influenza A(H1N1)pdm09 infection

“…Samuel et al (2011) generated mAbs to the HA1 of the H1N1pdm09 viral HA for the serodiagnosis of H1N1-pdm09 infections. Using an anti-NP mAb generated by the hybridoma technique with the immunization of NP that originated from the H5N1 virus, an immunochromatographic assay specific to the H1N1pdm09 virus was generated by another group (Miyoshi-Akiyama et al, 2010). Most of the mAbs in these studies are insufficiently characterized except for their specificities to the H1N1pdm09 virion by HAI, ELISA, or Western blot.…”