Poster: Diseases of Plants: Disease Detection & Diagnosis
Discriminating Potexvirus species by RT-PCR coupled to High Resolution Melting
A. OLMEDO-VELARDE (1), F. Ochoa-Corona (2) (1) Universidad de las Fuerzas Armadas ESPE, Ecuador; (2) Oklahoma State University, U.S.A.
Potexvirus is a cosmopolitan plant virus genus and most of its thirty-seven confirmed species are mechanically transmitted, causing mosaic and ringspot symptoms on herbaceous hosts. High Resolution Melting analysis (HRM) allows the discrimination at species and strain levels and has application in diagnostics. Complete genome sequences for Alternanthera mosaic virus (AltMV), Clover yellow mosaic virus (ClYMV), Lettuce virus X (LeVX), Opuntia virus X (OpVX), Papaya mosaic virus (PapMV), Pepino mosaic virus (PepMV), Potato aucuba mosaic virus (PAMV), Potato virus X (PVX), Schlumbergera virus X (SchVX), Strawberry mild yellow edge virus (SMYEV) and Hosta virus X (HVX), were retrieved from NCBI Genebank and aligned using Mega 6 and screened for conserved domains. Primer set Potex F3/R3 was selected to discriminate among eleven potexviruses by a melting temperature prediction using uMeltSM, a melting curve prediction software program. Reference positive controls for these eleven viruses were obtained from DMSZ (Germany) and Agdia, Inc. (USA) and tested using RT-PCR coupled to HRM with Potex F3/R3. All viruses tested positive and showed different melting temperatures ranging from 77.9°C to 82.4°C allowing for their discrimination through HRM analysis. The combination of RT-PCR-HRM can be used in diagnostic laboratories to discriminate eleven Potexviruses with high confidence.