title: A new method for detection and discrimination of Pepino mosaic virus isolates using high resolution melting analysis of the triple gene block 3.
abstract: Diagnostic methods distinguished different Pepino mosaic virus (PepMV) genotypes but the methods do not detect sequence variation in particular gene segments. The necrotic and non-necrotic isolates (pathotypes) of PepMV share a 99% sequence similarity. These isolates differ from each other at one nucleotide site in the triple gene block 3. In this study, a combination of real-time reverse transcription polymerase chain reaction and high resolution melting curve analysis of triple gene block 3 was developed for simultaneous detection and differentiation of PepMV pathotypes. The triple gene block 3 region carrying a transition A → G was amplified using two primer pairs from twelve virus isolates, and was subjected to high resolution melting curve analysis. The results showed two distinct melting curve profiles related to each pathotype. The results also indicated that the high resolution melting method could readily differentiate between necrotic and non-necrotic PepMV pathotypes.
authors.affiliation: Institute of Plant Protection - National Research Institute, Department of Virology and Bacteriology, Władysława Węgorka 20, 60-318 Poznań, Poland. B.Hasiow@iorpib.poznan.pl
authors.forename: Beata
authors.initials: B
authors.lastname: Hasiów-Jaroszewska
authors.forename: Beata
authors.initials: B
authors.lastname: Komorowska
mesh_terms: D025202:Molecular Diagnostic Techniques / Q000379:methods*; D017354:Point Mutation; D017863:Potexvirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D044366:Transition Temperature; D014764:Viral Proteins / Q000235:genetics*; D014773:Virology / Q000379:methods*
chemical_list: D014764:Viral Proteins
keywords: Detection; HRM; PepMV; RT-PCR; Triple gene block 3