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Article Abstract
Detection of neutralizing antibody to viral neuraminidase (NA) by testing for enzyme inhibition has been recognized as an important part of the immunogenicity of influenza vaccines. However, the absence of a well characterized standard source of active NA and validated assays has significantly limited clinical studies of NA immunity. Influenza virus-like particles (VLPs) containing hemagglutinin (HA), NA, and M1 proteins were produced from insect cells infected with a recombinant baculovirus and used as the NA source for the NA inhibition (NAI) assay. The NA activity of 6 different VLP strains varied from 0.43 to 1.61 (×10(-3)) enzyme units per μg of HA and was stable over 6 months of storage at 2-8°C. The NAI assay using 2'-(4-methylumbelliferyl)-α-d-N-acetylneuraminic acid as a substrate was modified for testing the antibody titer in clinical samples and validated. The advantages of the assay include: (1) stable, reproducible, and standardized source of NA; (2) testing the antibody titer specific to each subtype of NA in serum from subjects immunized with trivalent vaccines (H1N1, H3N2, B) with no interference from antibodies specific to the HA and to heterologous subtypes of the NA; (3) suitability for conducting long-term clinical trials as a result of low intra- and inter-assay variability, and (4) a wide analytical range due to 25% inhibition cut-off value for the NAI titer estimation.
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| http://dx.doi.org/10.1016/j.jviromet.2011.03.011 | DOI Listing |
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