基于重组VP9的酶联免疫吸附测定法检测针对版纳病毒（赛多病毒属）的免疫球蛋白G抗体

Recombinant VP9-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Banna virus (genus Seadornavirus).

作者信息

Mohd Jaafar Fauziah, Attoui Houssam, Gallian Pierre, Isahak Ilina, Wong Kum Thong, Cheong Soon Keng, Nadarajah Veera Sekaran, Cantaloube Jean-François, Biagini Philippe, De Micco Philippe, De Lamballerie Xavier

机构信息

Unité des Virus Emergents (EA 3292, IFR 48): Université de la Méditerranée, Faculté de Médecine de Marseille and EFS Alpes-Méditerranée, Marseille 13005, France.

出版信息

J Virol Methods. 2004 Mar 1;116(1):55-61. doi: 10.1016/j.jviromet.2003.10.010.

DOI:10.1016/j.jviromet.2003.10.010

PMID:14715307

Abstract

Banna virus (BAV, genus Seadornavirus, family Reoviridae) is an arbovirus suspected to be responsible for encephalitis in humans. Two genotypes of this virus are distinguishable: A (Chinese isolate, BAV-Ch) and B (Indonesian isolate, BAV-In6969) which exhibit only 41% amino-acid identity in the sequence of their VP9. The VP7 to VP12 of BAV-Ch and VP9 of BAV-In6969 were expressed in bacteria using pGEX-4T-2 vector. VP9 was chosen to establish an ELISA for BAV, based mainly on two observations: (i). VP9 is a major protein in virus-infected cells and is a capsid protein (ii). among all the proteins expressed, VP9 was obtained in high amount and showed the highest immuno-reactivity to anti-BAV ascitic fluid. The VP9s ELISA was evaluated in three populations: French blood donors and two populations (blood donors and patients with a neurological syndrome) from Malaysia, representing the region where the virus was isolated in the past. The specificity of this ELISA was >98%. In mice injected with live BAV, the assay detected IgG-antibody to BAV infection 21 days post-injection, which was confirmed by Western blot using BAV-infected cells. The VP9 ELISA permits to determine the sero-status of a population without special safety precautions and without any requirements to propagate the BAV. This test should be a useful tool for epidemiological survey of BAV.

摘要

版纳病毒（BAV，属于呼肠孤病毒科海多病毒属）是一种虫媒病毒，被怀疑是导致人类脑炎的病原体。该病毒有两种基因型可区分：A（中国分离株，BAV-Ch）和B（印度尼西亚分离株，BAV-In6969），它们的VP9序列氨基酸同一性仅为41%。利用pGEX-4T-2载体在细菌中表达了BAV-Ch的VP7至VP12以及BAV-In6969的VP9。选择VP9建立BAV的ELISA检测方法，主要基于两点观察：（i）VP9是病毒感染细胞中的主要蛋白，是一种衣壳蛋白；（ii）在所有表达的蛋白中，VP9产量高，且对抗BAV腹水的免疫反应性最强。在三个群体中对VP9 ELISA进行了评估：法国献血者以及来自马来西亚的两个群体（献血者和患有神经综合征的患者），马来西亚代表过去分离出该病毒的地区。该ELISA的特异性>98%。在注射活BAV的小鼠中，该检测方法在注射后21天检测到针对BAV感染的IgG抗体，这通过使用BAV感染细胞的蛋白质印迹法得到证实。VP9 ELISA无需特殊安全防护措施，也无需增殖BAV，就能确定群体的血清状态。该检测方法应是BAV流行病学调查的有用工具。