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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 PublicationJOURNAL OF VIROLOGICAL METHODS
ISSN0166-0934
Volume169Issue:1Pages:28-33
AbstractAn 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.
KeywordDsrna 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 AreaBiochemical Research Methods ; Biotechnology & Applied Microbiology ; Virology
Funding OrganizationNational 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 BySCI
Language英语
WOS IDWOS:000282398600005
Funding OrganizationNational 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]
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Cited Times:43[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://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 AuthorFang, 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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