A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus

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	A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus
	Yuan, Li 1; Zhang, Xiaohua 1; Chang, Mingxian 1; Jia, Chensong 1; Hemmingsen, Sean M.2; Dai, Heping 1
刊名	JOURNAL OF VIROLOGICAL METHODS
	2007-12-01
卷号	146 期号:1-2 页码:96-103
关键词	white spot syndrome virus (WSSV) fluorescent quantitative PCR neutralization assay primary cell culture
ISSN号	0166-0934
通讯作者	Dai, HP, Chinese Acad Sci, Inst Hydrobiol, E Lake So Rd 7, Wuhan 430072, Hubei, Peoples R China
中文摘要	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.
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.
学科主题	Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology
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
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
收录类别	SCI
语种	英语
WOS记录号	WOS:000251648400013
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/8334]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
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
推荐引用方式 GB/T 7714	Yuan, Li,Zhang, Xiaohua,Chang, Mingxian,et al. A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus[J]. JOURNAL OF VIROLOGICAL METHODS,2007,146(1-2):96-103.
APA	Yuan, Li,Zhang, Xiaohua,Chang, Mingxian,Jia, Chensong,Hemmingsen, Sean M.,&Dai, Heping.(2007).A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus.JOURNAL OF VIROLOGICAL METHODS,146(1-2),96-103.
MLA	Yuan, Li,et al."A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus".JOURNAL OF VIROLOGICAL METHODS 146.1-2(2007):96-103.