Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: Sensitive and high-throughput analysis utilizing secreted alkaline phosphatase

被引：37

作者：

Kaku, Yoshihiro ^{[1
]}

Noguchi, Akira ^{[1
]}

Marsh, Glenn A. ^{[2
]}

Barr, Jennifer A. ^{[2
]}

Okutani, Akiko ^{[1
]}

Hotta, Kozue ^{[1
]}

Bazartseren, Boldbaatar ^{[1
]}

Fukushi, Shuetsu ^{[3
]}

Broder, Christopher C. ^{[4
]}

Yamada, Akio ^{[1
]}

Inoue, Satoshi ^{[1
]}

Wang, Lin-Fa ^{[2
]}

机构：

[1] Natl Inst Infect Dis, Dept Vet Sci, Shinjuku Ku, Tokyo 1628640, Japan

[2] CSIRO Livestock Ind, Australian Anim Hlth Lab, Geelong, Vic 3220, Australia

[3] Natl Inst Infect Dis, Dept Virol 1, Tokyo 2080011, Japan

[4] Uniformed Serv Univ Hlth Sci, Bethesda, MD 20814 USA

来源：

JOURNAL OF VIROLOGICAL METHODS | 2012年 / 179卷 / 01期

关键词：

Henipavirus; Pseudotyped virus; Secreted alkaline phosphatase; Neutralization; Biosafety level 4; TO-PERSON TRANSMISSION; BATS; HENIPAVIRUS; BANGLADESH; INFECTION; OUTBREAK; HENDRA;

D O I：

10.1016/j.jviromet.2011.11.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra virus (Hey). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped with NiV F/G proteins (VSV-NiV-SEAP). A unique characteristic of this novel assay is the ability to obtain neutralization titers by measuring SEAP activity in supernatant using a common ELISA plate reader. This confers a remarkable advantage over the first generation of NiV-pseudotypes expressing green fluorescent protein or luciferase, which require expensive and specific measuring equipment. Using panels of NiV- and Hey-specific sera from various species, the VSV-NiV-SEAP assay demonstrated neutralizing antibody status (positive/negative) consistent with that obtained by conventional live NiV test, and gave higher antibody titers than the latter. Additionally, when screening sixty-six fruit bat sera at one dilution, the VSV-NiV-SEAP assay produced identical results to the live NiV test and only required a very small amount (2 mu l) of sera. The results suggest that this novel VSV-NiV-SEAP assay is safe, useful for high-throughput screening of sera using an ELISA plate reader, and has high sensitivity and specificity. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：226 / 232

页数：7

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