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Rapid detection of potyviruses from crude plant extracts.

Authors
  • Silva, Gonçalo1
  • Oyekanmi, Joshua2
  • Nkere, Chukwuemeka K3
  • Bömer, Moritz4
  • Kumar, P Lava2
  • Seal, Susan E4
  • 1 Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent, ME4 4TB, UK. Electronic address: [email protected]
  • 2 International Institute of Tropical Agriculture (IITA), Oyo Road, PMB 5320, Ibadan, Nigeria. , (Niger)
  • 3 International Institute of Tropical Agriculture (IITA), Oyo Road, PMB 5320, Ibadan, Nigeria; National Root Crops Research Institute, Km 8 Ikot Ekpene Road, PMB 7006, Umudike, Nigeria. , (Niger)
  • 4 Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent, ME4 4TB, UK.
Type
Published Article
Journal
Analytical Biochemistry
Publisher
Elsevier
Publication Date
Jan 31, 2018
Volume
546
Pages
17–22
Identifiers
DOI: 10.1016/j.ab.2018.01.019
PMID: 29378167
Source
Medline
Keywords
License
Unknown

Abstract

Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particularly in plant species containing high amounts of polysaccharides and polyphenols such as yam (Dioscorea spp.). Here, we report the development of a reliable, rapid and cost-effective detection method for the two most important potyviruses infecting yam based on reverse transcription-recombinase polymerase amplification (RT-RPA). The developed method, named 'Direct RT-RPA', detects each target virus directly from plant leaf extracts prepared with a simple and inexpensive extraction method avoiding laborious extraction of high-quality RNA. Direct RT-RPA enables the detection of virus-positive samples in under 30 min at a single low operation temperature (37 °C) without the need for any expensive instrumentation. The Direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide.

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