A neutralization test for specific detection of Nipah virus antibodies using pseudotyped vesicular stomatitis virus expressing green fluorescent protein

• Published in: Journal of virological methods Vol. 160; no. 1; pp. 7 - 13
• Main Authors: Kaku, Yoshihiro, Noguchi, Akira, Marsh, Glenn A., McEachern, Jennifer A., Okutani, Akiko, Hotta, Kozue, Bazartseren, Boldbaatar, Fukushi, Shuetsu, Broder, Christopher C., Yamada, Akio, Inoue, Satoshi, Wang, Lin-Fa
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01-09-2009; Elsevier; Elsevier B.V. Published by Elsevier B.V
• Subjects: Animals; Antibodies, Viral - blood; Antigens, Viral - genetics; Biological and medical sciences; Biosafety; Cats; Chiroptera; Diagnosis; Fundamental and applied biological sciences. Psychology; Genes, Reporter; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Hendra virus; Hendra Virus - immunology; Henipavirus; Henipavirus Infections - diagnosis; Henipavirus Infections - immunology; Hepatitis E virus; Horses; Humans; Microbiology; Neutralization; Neutralization Tests - methods; Nipah; Nipah virus; Nipah Virus - immunology; Paramyxovirus; Pseudotype; Rabbits; Sensitivity and Specificity; Swine; Techniques used in virology; Vesicular stomatitis virus; Vesiculovirus - genetics; Viral Proteins - genetics; Virology; Nipah; Pseudotype; Henipavirus; Diagnosis; Biosafety; Neutralization; Microbiology; Antibody; Method; Virology; Virus; Vesicular stomatitis virus; Mononegavirales; Vesiculovirus; Neutralization test; Rhabdoviridae; Green fluorescent protein; Nipah virus; Detection
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2009.04.037

Nipah virus (NiV) is a new zoonotic paramyxovirus that emerged in 1998 and is now classified in the genus
Henipavirus along with the closely related Hendra virus (HeV). NiV is highly pathogenic in several vertebrate species including humans, and the lack of available vaccines or specific treatment restricts it to biosafety level 4 (BSL4) containment. A serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing green fluorescent protein (GFP) and bearing the F and G proteins of NiV (VSV–NiV–GFP). The neutralization titers were obtained by counting GFP-expressing cells or by measuring fluorescence. The performance of this new assay was compared against the conventional test using live NiV with panels of sera from several mammalian species, including sera from NiV outbreaks, experimental infections, as well as HeV-specific sera. The results obtained with the VSV–NiV–GFP based test correlated with those obtained using live NiV. Using a 50% reduction in VSV–NiV–GFP infected cells as the cut-off for neutralization, this new assay demonstrated its potential as an effective tool for detecting NiV neutralizing antibodies under BSL2 containment with greater speed, sensitivity and safety as compared to the conventional NiV serum neutralization test.