Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: sensitive and high-throughput analysis utilizing secreted alkaline phosphatase.
作者： Yoshihiro Kaku , Akira Noguchi , Glenn A. Marsh , Jennifer A. Barr , Akiko Okutani
DOI: 10.1016/J.JVIROMET.2011.11.003
关键词: Neutralization 、 Vesicular stomatitis virus 、 Blood serum 、 Henipavirus 、 Titer 、 Biology 、 Neutralizing antibody 、 Paramyxoviridae 、 Antibody titer 、 Virology
摘要: Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra (HeV). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using recombinant vesicular stomatitis (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped F/G proteins (VSV-NiV-SEAP). unique characteristic of this assay ability to obtain titers by SEAP activity in supernatant common ELISA plate reader. This confers remarkable advantage over first generation NiV-pseudotypes green fluorescent protein or luciferase, which require expensive specific equipment. Using panels NiV- HeV-specific sera from various species, VSV-NiV-SEAP demonstrated antibody status (positive/negative) consistent that obtained conventional live test, gave higher than latter. Additionally, when screening sixty-six fruit bat at one dilution, produced identical results only required very small amount (2μl) sera. The suggest safe, useful high-throughput an reader, has high sensitivity specificity.
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