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      Safe Pseudovirus-based Assay for Neutralization Antibodies against Influenza A(H7N9) Virus

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          Abstract

          Serologic studies are urgently needed to assist in understanding an outbreak of influenza A(H7N9) virus. However, a biosafety level 3 laboratory is required for conventional serologic assays with live lethal virus. We describe a safe pseudovirus–based neutralization assay with preliminary assessment using subtype H7N9–infected samples and controls.

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          Human Infection with a Novel Avian-Origin Influenza A (H7N9) Virus

          New England Journal of Medicine, 368(20), 1888-1897
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            Hemagglutinin pseudotyped lentiviral particles: characterization of a new method for avian H5N1 influenza sero-diagnosis.

            Highly pathogenic avian influenza (HPAI) H5N1 has spread globally in birds and infected over 270 humans with an apparently high mortality rate. Serologic studies to determine the extent of asymptomatic H5N1 infection in humans and other mammals and to investigate the immunogenicity of current H5N1 vaccine candidates have been hampered by the biosafety requirements needed for H5N1 micro-neutralization tests. Development of a serodiagnostic tool for highly pathogenic influenza that reproduces H5N1 biology but can be used with less biohazard. We have generated and evaluated H5 hemagglutinin pseudotyped lentiviral particles encoding the luciferase reporter (H5pp). H5pp entry into target cells depends on alpha2-3 cell surface sialic acids and requires low pH for membrane fusion. H5pp infectivity is specifically neutralized by sera from patients and animals infected with H5N1 and correlates well with conventional microneutralization test. H5pp reproduce H5N1 influenza virus entry into target cells and potentially provides a high-throughput and safe method for sero-epidemiology.
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              Detection of Extensive Cross-Neutralization between Pandemic and Seasonal A/H1N1 Influenza Viruses Using a Pseudotype Neutralization Assay

              Background Cross-immunity between seasonal and pandemic A/H1N1 influenza viruses remains uncertain. In particular, the extent that previous infection or vaccination by seasonal A/H1N1 viruses can elicit protective immunity against pandemic A/H1N1 is unclear. Methodology/Principal Findings Neutralizing titers against seasonal A/H1N1 (A/Brisbane/59/2007) and against pandemic A/H1N1 (A/California/04/2009) were measured using an HIV-1-based pseudovirus neutralization assay. Using this highly sensitive assay, we found that a large fraction of subjects who had never been exposed to pandemic A/H1N1 express high levels of pandemic A/H1N1 neutralizing titers. A significant correlation was seen between neutralization of pandemic A/H1N1 and neutralization of a standard seasonal A/H1N1 strain. Significantly higher pandemic A/H1N1 neutralizing titers were measured in subjects who had received vaccination against seasonal influenza in 2008–2009. Higher pandemic neutralizing titers were also measured in subjects over 60 years of age. Conclusions/Significance Our findings reveal that the extent of protective cross-immunity between seasonal and pandemic A/H1N1 influenza viruses may be more important than previously estimated. This cross-immunity could provide a possible explanation of the relatively mild profile of the recent influenza pandemic.
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                Author and article information

                Journal
                Emerg Infect Dis
                Emerging Infect. Dis
                EID
                Emerging Infectious Diseases
                Centers for Disease Control and Prevention
                1080-6040
                1080-6059
                October 2013
                : 19
                : 10
                : 1685-1687
                Affiliations
                [1]Shanghai Public Health Clinical Center, Shanghai, China (C. Qiu, A. Zhang, D. Tian, Y. Wan, Xiaoling Zhang, W. Zhang, Z. Zhang, Z. Yuan, Y. Hu, Xiaoyan Zhang, J. Xu);
                [2]Fudan University, Shanghai (C. Qiu, Y. Huang, A. Zhang, Z. Yuan, Xiaoyan Zhang, J. Xu);
                [3]Chinese Center for Disease Control and Prevention, Beijing, China (Xiaoyan Zhang, J. Xu)
                Author notes
                Address for correspondence: Jianqing Xu, Shanghai Public Health Clinical Center, No. 2901, Caolang Rd, Jinshan District, Shanghai, Shanghai 201508, China; email: jianqingxu2008@ 123456gmail.com
                Article
                13-0728
                10.3201/eid1910.130728
                3810762
                24047684
                0351adc7-75f4-4be1-9234-863a3bac0c7a
                Categories
                Dispatch
                Dispatch

                Infectious disease & Microbiology
                influenza,subtype h7n9,serology,neutralization tests,pseudovirus,viruses

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