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学科主题: Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology
题名: A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus
作者: Yuan, Li1; Zhang, Xiaohua1; Chang, Mingxian1; Jia, Chensong1; Hemmingsen, Sean M.2; Dai, Heping1
通讯作者: Dai, HP, Chinese Acad Sci, Inst Hydrobiol, E Lake So Rd 7, Wuhan 430072, Hubei, Peoples R China
关键词: white spot syndrome virus (WSSV) ; fluorescent quantitative PCR ; neutralization assay ; primary cell culture
刊名: JOURNAL OF VIROLOGICAL METHODS
发表日期: 2007-12-01
DOI: 10.1016/j.jviromet.2007.06.009
卷: 146, 期:1-2, 页:96-103
收录类别: SCI
文章类型: Article
部门归属: [Yuan, Li; Zhang, Xiaohua; Chang, Mingxian; Jia, Chensong; Dai, Heping] Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China; [Hemmingsen, Sean M.] Natl Res Council Canada, Inst Plant Biotechnol, Saskatoon, SK S7N 0W9, Canada
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology
摘要: A fluorescent quantitative PCR (FQ-PCR) assay utilizing SYBR green I dye is described for quantitation of white spot syndrome virus (WSSV) particles isolated from infected crayfish, Cambarus clarkii. For this assay, a primer set was designed which amplifies, with high efficiency and specificity, a 129 bp target sequence within ORF167 of the WSSV genome. Conveniently, WSSV particles can be added into the FQ-PCR assay with a simple and convenient method to release its DNA. To establish the basis for an in vitro neutralization test, primary cultures of shrimp cells were challenged with WSSV that had been incubated with a polyclonal anti-WSSV serum or with control proteins. The number of WSSV particles released from the cells after these treatments were assayed by FQ-PCR. This test may serve as a method to screen monoclonal antibody pools or recombinant antibody pools for neutralizing activity prior to in vivo animal experiments. (c) 2007 Elsevier B.V. All rights reserved.
英文摘要: A fluorescent quantitative PCR (FQ-PCR) assay utilizing SYBR green I dye is described for quantitation of white spot syndrome virus (WSSV) particles isolated from infected crayfish, Cambarus clarkii. For this assay, a primer set was designed which amplifies, with high efficiency and specificity, a 129 bp target sequence within ORF167 of the WSSV genome. Conveniently, WSSV particles can be added into the FQ-PCR assay with a simple and convenient method to release its DNA. To establish the basis for an in vitro neutralization test, primary cultures of shrimp cells were challenged with WSSV that had been incubated with a polyclonal anti-WSSV serum or with control proteins. The number of WSSV particles released from the cells after these treatments were assayed by FQ-PCR. This test may serve as a method to screen monoclonal antibody pools or recombinant antibody pools for neutralizing activity prior to in vivo animal experiments. (c) 2007 Elsevier B.V. All rights reserved.
关键词[WOS]: POLYMERASE-CHAIN-REACTION ; PASSIVE LATEX AGGLUTINATION ; CULTURED PENAEID SHRIMP ; ENVELOPE PROTEIN VP28 ; SITU HYBRIDIZATION ; BACULOVIRUS WSBV ; MONOCLONAL-ANTIBODIES ; INFECTION ; WSSV ; MONODON
语种: 英语
WOS记录号: WOS:000251648400013
ISSN号: 0166-0934
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/8334
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
2.Natl Res Council Canada, Inst Plant Biotechnol, Saskatoon, SK S7N 0W9, Canada

Recommended Citation:
Yuan, Li; Zhang, Xiaohua; Chang, Mingxian; Jia, Chensong; Hemmingsen, Sean M.; Dai, Heping.A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus,JOURNAL OF VIROLOGICAL METHODS,2007,146(1-2):96-103
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