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Title: Development of a rapid, sensitive and specific diagnostic assay for fish Aquareovirus based on RT-PCR
Authors: Seng, E.K.
Fang, Q.
Lam, T.J. 
Sin, Y.M. 
Keywords: Aquareovirus
Degenerate primers
Threadfin reovirus
Issue Date: 15-Jun-2004
Citation: Seng, E.K., Fang, Q., Lam, T.J., Sin, Y.M. (2004-06-15). Development of a rapid, sensitive and specific diagnostic assay for fish Aquareovirus based on RT-PCR. Journal of Virological Methods 118 (2) : 111-122. ScholarBank@NUS Repository.
Abstract: 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 450bp 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. © 2004 Elsevier B.V. All rights reserved.
Source Title: Journal of Virological Methods
ISSN: 01660934
DOI: 10.1016/j.jviromet.2004.01.023
Appears in Collections:Staff Publications

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