A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus
文献类型：期刊论文
| 作者 | Yuan, Li1; Zhang, Xiaohua1; Chang, Mingxian1; Jia, Chensong1; Hemmingsen, Sean M.2; Dai, Heping1 |
| 刊名 | JOURNAL OF VIROLOGICAL METHODS
 |
| 出版日期 | 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. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 学科主题 | Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology |
| 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 |
| 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. |
入库方式： OAI收割
来源：水生生物研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。