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Molecular detection of genotype II grass carp reovirus based on nucleic acid sequence-based amplification combined with enzyme-linked immunosorbent assay (NASBA-ELISA)

文献类型： 外文期刊

作者： Zeng, Weiwei 1 ; Yao, Wei; Wang, Yingying 1 ; Li, Yingying 1 ; Bermann, Sven M.; Ren, Yan 1 ; Shi, Cunbin 1 ; Song, 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Key Lab Fishery Drug Dev, Key Lab Aquat Anim Immune Technol,Minist Agr, Guangzhou 510380, Guangdong, Peoples R China

2.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Key Lab Fishery Drug Dev, Key Lab Aquat Anim Immune Technol,Minist Agr, Guangzhou 5103

关键词： Grass carp reovirus;GCRV II;NASBA-ELISA

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

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收录情况： SCI

摘要： Grass carp reovirus (GCRV) is the causative agent of the grass carp hemorrhagic disease that has resulted in severe economic losses in the grass carp (Ctenopharyngodon idella) farming industry in China. Early diagnosis and vaccine administration are important priorities for GCRV control. In this study, a nucleic acid sequence-based amplification with enzyme-linked immunosorbent assay (NASBA-ELISA) was developed for to detect genotype II GCRV (GCRV- II). Primers specifically targeting viral RNA genome segment 6 were utilized for amplification in an isothermal digoxigenin-labeling NASBA process, resulting in DIG-labeled RNA amplicons. The amplicons were hybridized to specific biotinylated DNA probes and the products were detected colorimetrically using horseradish peroxidase and a microplate reader. The new method is able to detect GCRV at 14 copies/mu L within 5 h and had a diagnostic sensitivity and a specificity of 100% when GCRV-II and non-target virus were tested. This NASBA-ELISA was evaluated using a panel of clinical samples (n =103) to demonstrate that it is a rapid, effective and sensitive method for GCRV detection in grass carp aquaculture. (C) 2017 Elsevier B.V. All rights reserved.

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[1]Development of indirect immunofluorescence assay for TCID50 measurement of grass carp reovirus genotype II without cytopathic effect onto cells. Wang, Qing,Xie, Hualiang,Zeng, Weiwei,Liu, Chun,Wu, Jiexing,Wang, Yingying,Li, Yingying,Xie, Hualiang,Wang, Linchuan,Bergmann, Sven M.. 2018

[2]Molecular analysis of grass carp reovirus HZ08 genome segments 1-3 and 5-6. Zhang, Chao,Wang, Qing,Shi, Cunbin,Zeng, Weiwei,Liu, Yongkui,Wu, Shuqin,Zhang, Chao,Liu, Yongkui. 2010

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[4]Prokaryotic expression and immunoassay of grass carp reovirus capsid VP6 protein. Zhou, Y.,Fan, Y. D.,Zeng, L. B.,Ma, J.. 2013

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