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J Virol Methods. 2015 Sep 15;222:138-44. doi: 10.1016/j.jviromet.2015.06.011. Epub 2015 Jun 23.

Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification.

Author information

1
Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent ME4 4TB, UK. Electronic address: g.silva@gre.ac.uk.
2
Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent ME4 4TB, UK.
3
International Institute of Tropical Agriculture (IITA), Oyo Road, PMB 5320, Ibadan, Nigeria.

Abstract

Yam mosaic virus (YMV; genus Potyvirus) is considered to cause the most economically important viral disease of yams (Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase amplification (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/μl of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37°C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratories.

KEYWORDS:

Diagnosis; Dioscorea spp.; Recombinase polymerase amplification; Yam; Yam mosaic virus

PMID:
26115609
DOI:
10.1016/j.jviromet.2015.06.011
[Indexed for MEDLINE]

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