Expression of recombinant HA1 protein for specific detection of influenza A/H1N1/2009 antibodies in human serum

Abstract

The hemagglutinin genes (HA1 subunit) from human and animal 2009 pandemic H1N1 virus isolates were expressed with a baculovirus vector. Recombinant HA1 (rHA1) protein-based ELISA was evaluated for detection of specific influenza A(H1N1)pdm09 antibodies in serum samples from vaccinated humans. It was found that rHA1 ELISA consistently differentiated between antibodies recognizing the seasonal influenza H1N1 and pdm09 viruses, with a concordance of 94% as compared to the hemagglutination inhibition test. This study suggests the utility of rHA1 ELISA in serosurveillance.

Expression and Characterization of HA1 Protein of Highly Pathogenic H5N1 Avian Influenza Virus for Use in a Serodiagnostic Assay

“…Antibodies to the HA1 protein are subtype specific, so that antibodies against one subtype will not typically react with another subtype. In a previous study (Luo et al., ), we also showed the utility of H1N1 rHA1 ELISA in differentiating the seasonal influenza H1N1 and swine‐origin 2009 H1N1 viruses. In this study, we also demonstrated that the rHA1‐based indirect ELISA appears to be more sensitive, specific and easy to perform for detection of HPIA H5 subtype‐specific antibodies in chicken sera and subtyping of AIV at a single serum dilution.…”

“…The successful cloning of cDNA, encoding the HA1 gene with a melittin signal sequence and replacing a natural signal sequence and a 6xHis tag at the C‐terminus, into a baculovirus expression vector has provided the opportunity to produce large quantities of purified AIV rHA1 protein for use in studying intracellular processing and glycosylation of viral envelope proteins, characterizing antigenic relationships, diagnostic testing and vaccine development. The lack of reactivity of the H5 rHA1 protein with the other 15 subtypes would make it potentially suitable for specific ELISA‐based detection of H5 infections in poultry similar to that demonstrated for the AIV H1N1 rHA1 protein (Luo et al., ). Furthermore, the recombinant HA1 showed specific binding with anti‐H5N1‐specific antibody on the infected cells surface, indicating the HA1 protein could also be used as a diagnostic reagent in the immunofluorescence assay.…”

“…When highly pathogenic or novel influenza strains are involved, specialized laboratories with high‐level containment facilities and specially trained personnel are required for safe implementation of virus culture and testing (Alvarez et al., ). The ELISA has been recognized as a suitable alternative for diagnosis and seroepidemiological surveillance due to its simplicity, high sensitivity and economy (Alvarez et al., ; Luo et al., ).…”

HA1-specific indirect ELISA for serological detection of canine influenza virus H3N2 infection in dogs

“…The HA1 domain of HA is also associated with virus-neutralizing activity (Shoji et al, 2008;Tonegawa et al, 2003) and several ELISAs have used the HA1 protein as the coating antigen (Luo et al, 2013;Shahsavandi et al, 2011). In this study, an indirect ELISA was successfully established and tested using sera from dogs infected with Korean canine H3N2 viruses.…”

Seroprevalence of Pandemic H1n1 Influenza Among Pregnant Women in Kerala, India

“…10 In this study, ELISA using a monoclonal antibody (mAb) against influenza A Haemagglutinin protein (HP) -A/California/07/2009 was used to detect antibodies against influenza. Where H1N1 antigen was used as positive control, H1N1 negative human serum as negative control and patient serum were diluted from 1:10 to 1:2560.…”