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A new fluorescent quantitative PCR-based in vitro neutralization assay for white spot syndrome virus
Yuan, Li1; Zhang, Xiaohua1; Chang, Mingxian1; Jia, Chensong1; Hemmingsen, Sean M.2; Dai, Heping1; Dai, HP, Chinese Acad Sci, Inst Hydrobiol, E Lake So Rd 7, Wuhan 430072, Hubei, Peoples R China
2007-12-01
Source PublicationJOURNAL OF VIROLOGICAL METHODS
ISSN0166-0934
Volume146Issue:1-2Pages:96-103
AbstractA 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.; 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.
SubtypeArticle
KeywordWhite Spot Syndrome Virus (Wssv) Fluorescent Quantitative Pcr Neutralization Assay Primary Cell Culture
Department[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
Subject AreaBiochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
DOI10.1016/j.jviromet.2007.06.009
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology
WOS SubjectBiochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
WOS IDWOS:000251648400013
WOS KeywordPOLYMERASE-CHAIN-REACTION ; PASSIVE LATEX AGGLUTINATION ; CULTURED PENAEID SHRIMP ; ENVELOPE PROTEIN VP28 ; SITU HYBRIDIZATION ; BACULOVIRUS WSBV ; MONOCLONAL-ANTIBODIES ; INFECTION ; WSSV ; MONODON
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8334
Collection期刊论文
Corresponding AuthorDai, HP, Chinese Acad Sci, Inst Hydrobiol, E Lake So Rd 7, Wuhan 430072, Hubei, Peoples R China
Affiliation1.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
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, HP, Chinese Acad Sci, Inst Hydrobiol, E Lake So Rd 7, Wuhan 430072, Hubei, Peoples R China.(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.
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