An improved RT-PCR assay for rapid and sensitive detection of grass carp reovirus
文献类型：期刊论文
| 作者 | Zhang, Lanlan1; Luo, Qing2; Fang, Qin1; Wang, Yaping2 |
| 刊名 | JOURNAL OF VIROLOGICAL METHODS
 |
| 出版日期 | 2010-10-01 |
| 卷号 | 169期号:1页码:28-33 |
| 关键词 | dsRNA virus Aquareovirus Grass carp reovirus RNA extraction RT-PCR rapid detection |
| ISSN号 | 0166-0934 |
| 通讯作者 | Fang, Q, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China |
| 中文摘要 | An improved simple, rapid and sensitive method for detecting grass carp reovirus (GCRV) based on RT-PCR was developed by combining an advanced RNA extraction technique and targeting segment 10 as a template. The results indicate that highly efficient RT-PCR amplification of GCRV genome segments can be obtained using column-extracted RNA as a template, which is suitable not only for full-length gene amplification up to a size of 1.5 kb, but also for partial genome detection. Moreover, by targeting the highly divergent segment 10, the sensitivity of RT-PCR detection is improved significantly; as little as 1.0 fg of the 515 bp S10 dsRNA can be detected by one-step RT-PCR amplification. Furthermore, this method exhibits good reproducibility and specificity, and no amplicons were observed when RNA fragments other than those from GCRV were used as templates. The entire detection process can be completed within 4-5 h from RNA extraction, much faster than methods reported previously. Overall, the improved detection technique may be applied for rapid diagnosis of GCRV or other dsRNA viruses. (c) 2010 Elsevier B.V. All rights reserved. |
| 英文摘要 | An improved simple, rapid and sensitive method for detecting grass carp reovirus (GCRV) based on RT-PCR was developed by combining an advanced RNA extraction technique and targeting segment 10 as a template. The results indicate that highly efficient RT-PCR amplification of GCRV genome segments can be obtained using column-extracted RNA as a template, which is suitable not only for full-length gene amplification up to a size of 1.5 kb, but also for partial genome detection. Moreover, by targeting the highly divergent segment 10, the sensitivity of RT-PCR detection is improved significantly; as little as 1.0 fg of the 515 bp S10 dsRNA can be detected by one-step RT-PCR amplification. Furthermore, this method exhibits good reproducibility and specificity, and no amplicons were observed when RNA fragments other than those from GCRV were used as templates. The entire detection process can be completed within 4-5 h from RNA extraction, much faster than methods reported previously. Overall, the improved detection technique may be applied for rapid diagnosis of GCRV or other dsRNA viruses. (c) 2010 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 ; GENUS AQUAREOVIRUS ; FAMILY REOVIRIDAE ; DIAGNOSTIC ASSAY ; FISH ; VIRUS ; HYBRIDIZATION ; IDENTIFICATION ; GENOGROUP ; RNA |
| 收录类别 | SCI |
| 资助信息 | National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] |
| 语种 | 英语 |
| WOS记录号 | WOS:000282398600005 |
| 公开日期 | 2010-12-23 |
| 源URL | [http://ir.ihb.ac.cn/handle/342005/13579]  |
| 专题 | 水生生物研究所_鱼类生物学及渔业生物技术研究中心_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China 2.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Lanlan,Luo, Qing,Fang, Qin,et al. An improved RT-PCR assay for rapid and sensitive detection of grass carp reovirus[J]. JOURNAL OF VIROLOGICAL METHODS,2010,169(1):28-33. |
| APA | Zhang, Lanlan,Luo, Qing,Fang, Qin,&Wang, Yaping.(2010).An improved RT-PCR assay for rapid and sensitive detection of grass carp reovirus.JOURNAL OF VIROLOGICAL METHODS,169(1),28-33. |
| MLA | Zhang, Lanlan,et al."An improved RT-PCR assay for rapid and sensitive detection of grass carp reovirus".JOURNAL OF VIROLOGICAL METHODS 169.1(2010):28-33. |
入库方式： OAI收割
来源：水生生物研究所
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