| 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 Publication | JOURNAL OF VIROLOGICAL METHODS
 |
| ISSN | 0166-0934 |
| Volume | 146Issue:1-2Pages:96-103 |
| Abstract | 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.; 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. |
| Subtype | Article |
| Keyword | White 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 Area | Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology |
| DOI | 10.1016/j.jviromet.2007.06.009 |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| Indexed By | SCI |
| Language | 英语 |
| WOS Research Area | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Virology |
| WOS Subject | Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology |
| WOS ID | WOS:000251648400013 |
| WOS Keyword | POLYMERASE-CHAIN-REACTION ; PASSIVE LATEX AGGLUTINATION ; CULTURED PENAEID SHRIMP ; ENVELOPE PROTEIN VP28 ; SITU HYBRIDIZATION ; BACULOVIRUS WSBV ; MONOCLONAL-ANTIBODIES ; INFECTION ; WSSV ; MONODON |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ihb.ac.cn/handle/152342/8334 |
| Collection | 期刊论文 |
| Corresponding Author | Dai, HP, Chinese Acad Sci, Inst Hydrobiol, E Lake So Rd 7, Wuhan 430072, Hubei, Peoples R China |
| Affiliation | 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 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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