Development of a rapid, sensitive and specific diagnostic assay for fish Aquareovirus based on RT-PCR
文献类型：期刊论文
| 作者 | Seng, EK; Fang, Q; Lam, TJ; Sin, YM |
| 刊名 | JOURNAL OF VIROLOGICAL METHODS
 |
| 出版日期 | 2004-06-15 |
| 卷号 | 118期号:2页码:111-122 |
| ISSN号 | 0166-0934 |
| 关键词 | Aquareovirus RT-PCR degenerate primers threadfin reovirus |
| 通讯作者 | Seng, EK, Natl Univ Singapore, Dept Biol Sci, Block S1A,05-02 Virol Lab,14 Sci Dr 4, Singapore 117543, Singapore |
| 中文摘要 | A rapid, sensitive and highly specific detection method for Aquareovirus based on reverse-transcription polymerase chain reaction (RT-PCR) was developed. Based on multiple sequence alignment of the cloned sequences of a local isolates, the Threadfin reovirus (TFV) and Guppy reovirus (GPV) with Grass carp reovirus (GCRV), a pair of degenerate primers was selected carefully and synthesized. Using this primer combination, only one specific product, approximately 450 bp in length was obtained when RT-PCR was carried out using the genomic double-stranded RNA (dsRNA) of TFV, GPV and GCRV. Similar results were also obtained when Chum salmon reovirus (CSRV) and Striped bass reovirus (SBRV) dsRNA were used as templates. No products were observed when nucleic acids other than the dsRNA of the aquareoviruses described above were used as RT-PCR templates. This technique could detect not only TFV but also GPV and GCRV in low titer virus-infected cell cultured cells. Furthermore, this method has also been shown to be able to diagnose GPV-infected guppy (Poecilia reticulata) that exhibit clinical symptoms as well as GPV-carrier guppy. Collectively, these results showed that the RT-PCR amplification method using specific degenerate primers described below is very useful for rapid and accurate detection of a variety of aquareovirus strains isolated from different host species and origin. (C) 2004 Elsevier B.V. All rights reserved. |
| 英文摘要 | A rapid, sensitive and highly specific detection method for Aquareovirus based on reverse-transcription polymerase chain reaction (RT-PCR) was developed. Based on multiple sequence alignment of the cloned sequences of a local isolates, the Threadfin reovirus (TFV) and Guppy reovirus (GPV) with Grass carp reovirus (GCRV), a pair of degenerate primers was selected carefully and synthesized. Using this primer combination, only one specific product, approximately 450 bp in length was obtained when RT-PCR was carried out using the genomic double-stranded RNA (dsRNA) of TFV, GPV and GCRV. Similar results were also obtained when Chum salmon reovirus (CSRV) and Striped bass reovirus (SBRV) dsRNA were used as templates. No products were observed when nucleic acids other than the dsRNA of the aquareoviruses described above were used as RT-PCR templates. This technique could detect not only TFV but also GPV and GCRV in low titer virus-infected cell cultured cells. Furthermore, this method has also been shown to be able to diagnose GPV-infected guppy (Poecilia reticulata) that exhibit clinical symptoms as well as GPV-carrier guppy. Collectively, these results showed that the RT-PCR amplification method using specific degenerate primers described below is very useful for rapid and accurate detection of a variety of aquareovirus strains isolated from different host species and origin. (C) 2004 Elsevier B.V. All rights reserved. |
| 学科主题 | Biochemical Research Methods; Biotechnology & Applied Microbiology; Virology |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 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] | RNA BLOT HYBRIDIZATION ; CELL-CULTURE ; IDENTIFICATION ; VIRUSES ; GENOGROUP ; ROTAVIRUS ; INFECTION ; REOVIRUS ; PROBE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000221207300005 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://ir.ihb.ac.cn/handle/152342/9496]  |
| 专题 | 水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文 |
| 作者单位 | 1.Natl Univ Singapore, Dept Biol Sci, Singapore 117543, Singapore 2.CAS, Wuhan Inst Virol, Inst Hydrobiol, Wuhan 430071, Hubei, Peoples R China 3.Teo Way Yong & Sons Pte Ltd, Singapore Fish Breeding & Immunizat Ctr, Singapore, Singapore |
| 推荐引用方式 GB/T 7714 | Seng, EK,Fang, Q,Lam, TJ,et al. Development of a rapid, sensitive and specific diagnostic assay for fish Aquareovirus based on RT-PCR[J]. JOURNAL OF VIROLOGICAL METHODS,2004,118(2):111-122. |
| APA | Seng, EK,Fang, Q,Lam, TJ,&Sin, YM.(2004).Development of a rapid, sensitive and specific diagnostic assay for fish Aquareovirus based on RT-PCR.JOURNAL OF VIROLOGICAL METHODS,118(2),111-122. |
| MLA | Seng, EK,et al."Development of a rapid, sensitive and specific diagnostic assay for fish Aquareovirus based on RT-PCR".JOURNAL OF VIROLOGICAL METHODS 118.2(2004):111-122. |
入库方式： OAI收割
来源：水生生物研究所
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