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A specific and sensitive antigen capture assay for NS1 protein quantitation in Japanese encephalitis virus infection |
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| Authors: | Li Y Z Counor D Lu P Liang G D Vu T Q H Phan T N Huynh T K L Sun G Grandadam M Butrapet S Lavergne J P Flamand M Yu Y X Solomon T Buchy P Deubel V |
| Institution: | a Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Shanghai Institute for Biological Sciences, 411 Hefei Road, Shanghai 200025, China b Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (China CDC), 155 Changbai Road Changping District, Beijing 102206, China c Institut Pasteur of Ho Chi Minh City, Ho Chi Minh City, 167 Pasteur Street, 8 Ward, 3 District, Ho Chi Minh City, Viet Nam d National Institute of Hygiene and Epidemiology, 1, Pho Yersin, Hanoi 10000, Viet Nam e Institut Pasteur, National Reference Center for Arboviruses, 25 Dr Roux Road, 25724 Paris cedex 15, France f University of Liverpool, Delby Street, Liverpool L69 3GA, UK g Institute of Biology and Chemistry of Proteins, 7, passage du Vercors, 69367 Lyon cedex 07, France h Institut Pasteur, Unit of Bunyaviridae, 25 Dr Roux Road, 75724 Paris cedex 15, France i National Institute for the Control of Pharmaceutical and Biological Products, Tiantan Xili 2# Chongwen Qu, Beijing 100050, China j Institut Pasteur in Cambodia, 5 Boulevard Monivong, Phnom Penh, Cambodia |
| Abstract: | Japanese encephalitis virus (JEV) is a human pathogenic, mosquito-borne flavivirus that is endemic/epidemic in Asia. JEV is rarely detected or isolated from blood or cerebrospinal fluid (CSF), and detection of IgM is generally diagnostic of the infection. The flavivirus nonstructural glycoprotein NS1 is released transiently during flavivirus replication. The aim of this study was to set up a quantitative JEV NS1 antigen capture assay. A soluble hexameric form of JEV NS1 protein was produced in a stable Drosophila S2 cell clone and purified from supernatant fluids. Two IgG1 monoclonal antibodies (MAbs) with high affinity against two different epitopes of JEV NS1 antigen were used to develop an antigen-capture assay with a limit of detection of 0.2 ng ml−1 NS1. Up to 1 μg ml−1 JEV NS1 protein was released in supernatants of mammalian cells infected with JEV but <10 ng ml−1 was released in sera of virus-infected mice before the onset of encephalitis and death. Moreover, NS1 protein was detected at low levels (<10 ng ml−1) in 23.8% of sera and in 10.5% of CSF of patients diagnosed as IgM-positive for JEV. This quantitative test of NS1 protein is proposed for highly specific diagnosis of acute infection with JEV genotypes I to IV. |
| Keywords: | Japanese encephalitis Flavivirus Genotype NS1 protein Monoclonal antibody Antigen-capture |
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