| An improved RT-PCR assay for rapid and sensitive detection of grass carp reovirus | |
| Zhang, Lanlan; Luo, Qing; Fang, Qin; Wang, Yaping; Fang, Q, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China | |
| 2010 | |
| Source Publication | JOURNAL OF VIROLOGICAL METHODS
 |
| ISSN | 0166-0934 |
| Volume | 169Issue:1Pages:28-33 |
| Abstract | 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. |
| Keyword | Dsrna Virus Aquareovirus Grass Carp Reovirus Rna Extraction Rt-pcr Rapid Detection |
| Department | [Zhang, Lanlan; Fang, Qin] Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China; [Luo, Qing; Wang, Yaping] Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China |
| Subject Area | Biochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology |
| Funding Organization | National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] ; National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] ; National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] ; National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] |
| Indexed By | SCI |
| Language | 英语 |
| WOS ID | WOS:000282398600005 |
| Funding Organization | National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] ; National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] ; National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] ; National Basic Research Program of China [2009CB118701]; National Natural Scientific Foundation of China [30871940, 30671615] |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ihb.ac.cn/handle/342005/13579 |
| Collection | 鱼类生物学及渔业生物技术研究中心_期刊论文 |
| Corresponding Author | Fang, Q, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China |
| Recommended Citation 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,&Fang, Q, Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China.(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. |
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