TY - JOUR
T1 - Detection and Differentiation of Potato Virus Y Strains by Melting Analysis of an Oligonucleotide Virus Probe
AU - Rotem, N.
AU - Shtein, C.
AU - Rosner, A.
AU - Levy, D.
AU - Rabinowitch, H. D.
N1 - Publisher Copyright:
© 2016, The Potato Association of America.
PY - 2016/12/1
Y1 - 2016/12/1
N2 - A novel simple, fast qualitative method for detection and differentiation of PVY strains in potatoes by the LightCycler technology is described. Fluorescent-labeled probe designed to contain variable degree of homology with a ‘target sequence’ of several known reference PVY strains was annealed to PCR products of these viruses followed by a graded melting analysis. The specific characteristics of the melting curves enable the detection, distinction and differentiation of each of following four known PVY strains O-FL, O-RB, N and NTN, in a single reaction obviating the need for size or nucleotide sequence analyses. In addition, it was demonstrated that virus extracts from plants infected with more than one strain can be resolved using this procedure. The melting-curves of extracts from 55 market size tubers harvested randomly from commercial fields and compared with those of the reference virus strains, revealed the presence of O-FL, O-RB and NTN strains of PVY.
AB - A novel simple, fast qualitative method for detection and differentiation of PVY strains in potatoes by the LightCycler technology is described. Fluorescent-labeled probe designed to contain variable degree of homology with a ‘target sequence’ of several known reference PVY strains was annealed to PCR products of these viruses followed by a graded melting analysis. The specific characteristics of the melting curves enable the detection, distinction and differentiation of each of following four known PVY strains O-FL, O-RB, N and NTN, in a single reaction obviating the need for size or nucleotide sequence analyses. In addition, it was demonstrated that virus extracts from plants infected with more than one strain can be resolved using this procedure. The melting-curves of extracts from 55 market size tubers harvested randomly from commercial fields and compared with those of the reference virus strains, revealed the presence of O-FL, O-RB and NTN strains of PVY.
KW - Genotyping
KW - Molecular analysis
KW - PCR
KW - PVY
KW - RT-PCR
UR - http://www.scopus.com/inward/record.url?scp=84981275138&partnerID=8YFLogxK
U2 - 10.1007/s12230-016-9531-7
DO - 10.1007/s12230-016-9531-7
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AN - SCOPUS:84981275138
SN - 1099-209X
VL - 93
SP - 620
EP - 625
JO - American Journal of Potato Research
JF - American Journal of Potato Research
IS - 6
ER -