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      A sensitive retroviral pseudotype assay for influenza H5N1‐neutralizing antibodies

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          Abstract

          Background  The World Health Organisation (WHO) recommended the development of simple, safe, sensitive and specific neutralization assays for avian influenza antibodies. We have used retroviral pseudotypes bearing influenza H5 hemagglutinin (HA) as safe, surrogate viruses for influenza neutralization assays which can be carried out at Biosafety Level 2.

          Results  Using our assay, sera from patients who had recovered from infection with influenza H5N1, and sera from animals experimentally immunized or infected with H5 tested positive for the presence of neutralizing antibodies to H5N1. Pseudotype neutralizing antibody titers were compared with titers obtained by hemagglutinin inhibition (HI) assays and microneutralization (MN) assays using live virus, and showed a high degree of correlation, sensitivity and specificity.

          Conclusions  The pseudotype neutralization assay is as sensitive as horse erythrocyte HI and MN for the detection of antibodies to H5N1. It is safer, and can be applied in a high‐throughput format for human and animal surveillance and for the evaluation of vaccines.

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          Most cited references26

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          In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector.

          D Trono, I Verma, L Naldini … (1996)
          A retroviral vector system based on the human immunodeficiency virus (HIV) was developed that, in contrast to a murine leukemia virus-based counterpart, transduced heterologous sequences into HeLa cells and rat fibroblasts blocked in the cell cycle, as well as into human primary macrophages. Additionally, the HIV vector could mediate stable in vivo gene transfer into terminally differentiated neurons. The ability of HIV-based viral vectors to deliver genes in vivo into nondividing cells could increase the applicability of retroviral vectors in human gene therapy.
          Article 0 comments Cited 413 times – based on 0 reviews     Review now
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            Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays.

            T. Rowe, R A Abernathy, J Hu-Primmer … (1999)
            From May to December 1997, 18 cases of mild to severe respiratory illness caused by avian influenza A (H5N1) viruses were identified in Hong Kong. The emergence of an avian virus in the human population prompted an epidemiological investigation to determine the extent of human-to-human transmission of the virus and risk factors associated with infection. The hemagglutination inhibition (HI) assay, the standard method for serologic detection of influenza virus infection in humans, has been shown to be less sensitive for the detection of antibodies induced by avian influenza viruses. Therefore, we developed a more sensitive microneutralization assay to detect antibodies to avian influenza in humans. Direct comparison of an HI assay and the microneutralization assay demonstrated that the latter was substantially more sensitive in detecting human antibodies to H5N1 virus in infected individuals. An H5-specific indirect enzyme-linked immunosorbent assay (ELISA) was also established to test children's sera. The sensitivity and specificity of the microneutralization assay were compared with those of an H5-specific indirect ELISA. When combined with a confirmatory H5-specific Western blot test, the specificities of both assays were improved. Maximum sensitivity (80%) and specificity (96%) for the detection of anti-H5 antibody in adults aged 18 to 59 years were achieved by using the microneutralization assay combined with Western blotting. Maximum sensitivity (100%) and specificity (100%) in detecting anti-H5 antibody in sera obtained from children less than 15 years of age were achieved by using ELISA combined with Western blotting. This new test algorithm is being used for the seroepidemiologic investigations of the avian H5N1 influenza outbreak.
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              Influenza: lessons from past pandemics, warnings from current incidents.

              Taisuke Horimoto, Yoshihiro Kawaoka (2005)
              Recent outbreaks of highly pathogenic avian influenza A virus infections (H5 and H7 subtypes) in poultry and in humans (through direct contact with infected birds) have had important economic repercussions and have raised concerns that a new influenza pandemic will occur in the near future. The eradication of pathogenic avian influenza viruses seems to be the most effective way to prevent influenza pandemics, although this strategy has not proven successful so far. Here, we review the molecular factors that contribute to the emergence of pandemic strains.
              Article 0 comments Cited 215 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Influenza Other Respir Viruses
                Journal ID (iso-abbrev): Influenza Other Respir Viruses
                Journal ID (doi): 10.1111/(ISSN)1750-2659
                Journal ID (publisher-id): IRV
                Title: Influenza and Other Respiratory Viruses
                Publisher: Blackwell Publishing Ltd (Oxford, UK )
                ISSN (Print): 1750-2640
                ISSN (Electronic): 1750-2659
                Publication date (Electronic): 26 July 2007
                Publication date (Print): May 2007
                Volume: 1
                Issue: 3 ( doiID: 10.1111/irv.2007.1.issue-3 )
                Pages: 105-112
                Affiliations
                [ 1 ]MRC/UCL Centre for Medical Molecular Virology, Division of Infection and Immunity, University College London, London, UK
                [ 2 ]Health Protection Agency, Centre for Infections, London, UK
                [ 3 ]National Institute for Biological Standards and Control, Hertfordshire, UK
                [ 4 ]Veterinary Laboratories Agency, Addlestone, Surrey, UK
                [ 5 ]Oxford University Clinical Research Unit and Hospital for Tropical Diseases, Ho Chi Minh City, Viet Nam.
                Author notes
                [*]Dr Nigel Temperton, MRC/UCL Centre for Medical Molecular Virology, Division of Infection and Immunity, University College London, 46 Cleveland Street, London, W1T 4JF, United Kingdom. E‐mail: nigel.temperton@ 123456ucl.ac.uk
                Article
                Publisher ID: IRV016
                DOI: 10.1111/j.1750-2659.2007.00016.x
                PMC ID: 4941878
                PubMed ID: 19453415
                SO-VID: 2356aef9-7f59-40b0-a484-9f7ec9364950
                History
                Page count
                Figures: 2, Tables: 4, Pages: 8
                Categories
                Subject: Original Articles
                Custom metadata
                source-schema-version-number 2.0
                cover-date May 2007
                details-of-publishers-convertor Converter:WILEY_ML3GV2_TO_NLMPMC version:4.6.9 mode:remove_FC converted:04.11.2015

                ScienceOpen disciplines: Infectious disease & Microbiology
                Keywords: h5n1,influenza,pseudotypes,serology,viral neutralization
                Data availability:
                ScienceOpen disciplines: Infectious disease & Microbiology
                Keywords: h5n1, influenza, pseudotypes, serology, viral neutralization

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