Version 1
: Received: 6 January 2017 / Approved: 6 January 2017 / Online: 6 January 2017 (10:21:52 CET)
How to cite:
Zhang, L.; Lu, J.; Li, J.; Tang, R.; Peng, H.; Meng, F.; Hu, Y. Establish and Apply a Safe and Convenient Pseudovirus-Based Assay to Detect Neutralizing Antibodies against Influenza A (H7N9) Virus. Preprints2017, 2017010033. https://doi.org/10.20944/preprints201701.0033.v1
Zhang, L.; Lu, J.; Li, J.; Tang, R.; Peng, H.; Meng, F.; Hu, Y. Establish and Apply a Safe and Convenient Pseudovirus-Based Assay to Detect Neutralizing Antibodies against Influenza A (H7N9) Virus. Preprints 2017, 2017010033. https://doi.org/10.20944/preprints201701.0033.v1
Zhang, L.; Lu, J.; Li, J.; Tang, R.; Peng, H.; Meng, F.; Hu, Y. Establish and Apply a Safe and Convenient Pseudovirus-Based Assay to Detect Neutralizing Antibodies against Influenza A (H7N9) Virus. Preprints2017, 2017010033. https://doi.org/10.20944/preprints201701.0033.v1
APA Style
Zhang, L., Lu, J., Li, J., Tang, R., Peng, H., Meng, F., & Hu, Y. (2017). Establish and Apply a Safe and Convenient Pseudovirus-Based Assay to Detect Neutralizing Antibodies against Influenza A (H7N9) Virus. Preprints. https://doi.org/10.20944/preprints201701.0033.v1
Chicago/Turabian Style
Zhang, L., Fanyue Meng and Yuemei Hu. 2017 "Establish and Apply a Safe and Convenient Pseudovirus-Based Assay to Detect Neutralizing Antibodies against Influenza A (H7N9) Virus" Preprints. https://doi.org/10.20944/preprints201701.0033.v1
Abstract
In March 2013, a novel avian influenza A H7N9 virus was emerged in China, which cause rapidly progressive pneumonia and with a high fatality rate. Serologic studies to evaluate neutralizing antibodies of infected patients and birds are invaluable tools for immunogenicity research of H7N9 and epidemiological investigation. Conventional neutralization assays are laborious and time-consuming which also hampered by biosafety requirement. In this study, We construct and produce pseudovirus bearing the full-length hemagglutinin (HA) of H7N9 virus in the Env-defective, luciferase-expressing HIV-1 backbone. The production of lentiviral pseudovirus was analysed by HA gene specific real-time reverse-transcription PCR, transmission electron microscopy (TEM), and Western Blot assay to prove the nucleic acid replication, the morphology of virus, and the expression of HA protein in pseudovirus. After that pseudovirus based inhibition assay was established to detect neutralizing antibodies of a panel of serum samples. Our results demonstrated that H7N9 pseudovirus which had single-cycle infection was generated. By comparing the neutralization antibody titers, pseudovirus based neutralization test could be recognized as an alternative of conventional microneutralization (MN). Hence, we conclude that it is possible to use pseudovirus inhibition assay to screen sera samples, as well as evaluate vaccine-induced neutralizing antibodies against H7N9 virus.
Keywords
H7N9 avian influenza; pseudovirus; neutralization assay; relative luminescence units (RLU)
Subject
Biology and Life Sciences, Virology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.