20933015
not annotated - annotated - LINNAEUS only
Detection and discrimination of members of the family Luteoviridae by real-time PCR and SYBR(R) GreenER(TM) melting curve analysis.
This study investigated the suitability of a two step real-time RT-PCR melting curve analysis as a tool for the detection and discrimination of nine species in the plant virus family Luteoviridae, being Soybean dwarf virus [SbDV], Bean leafroll virus [BLRV], Beet chlorosis virus [BChV], Beet mild yellowing virus [BMYV], Beet western yellows virus [BWYV], Cereal yellow dwarf virus-RPV [CYDV-RPV], Cucurbit aphid-borne yellows virus [CABYV], Potato leafroll virus [PLRV] and Turnip yellows virus [TuYV]. Melting temperature and shape of the melting peak were analysed for 68 bp and 148 bp coat protein gene amplicons using SYBR(R) GreenER(TM) fluorescent dye. Specific melting peaks with unique melting temperature were observed for the various species of the family Luteoviridae using the 68 bp amplicon, but not with the 148 bp amplicon. Due to the high variability of sequences for some members of this family, different melting temperatures were also observed between different isolates of the species CYDV-RPV and TuYV. Nevertheless, discrimination between species was achieved for SbDV, BLRV, BChV, BMYV, CABYV and either PLRV or BWYV. Melting curve analysis, in this study, is a faster and more discriminatory alternative to gel electrophoresis of end-point PCR products for the detection of Luteoviridae infection.
Ann file
T1 Species 337 356 Soybean dwarf virus
N1 Reference T1 Taxonomy:12049
T2 Species 358 362 SbDV
N2 Reference T2 Taxonomy:12049
T3 Species 365 384 Bean leafroll virus
N3 Reference T3 Taxonomy:12041
T4 Species 386 390 BLRV
N4 Reference T4 Taxonomy:12041
T5 Species 393 413 Beet chlorosis virus
N5 Reference T5 Taxonomy:131082
T6 Species 415 419 BChV
N6 Reference T6 Taxonomy:131082
T7 Species 422 447 Beet mild yellowing virus
N7 Reference T7 Taxonomy:156690
T8 Species 449 453 BMYV
N8 Reference T8 Taxonomy:156690
T9 Species 456 482 Beet western yellows virus
N9 Reference T9 Taxonomy:12042
T10 Species 484 488 BWYV
N10 Reference T10 Taxonomy:12042
T11 Species 491 520 Cereal yellow dwarf virus-RPV
T12 Species 522 530 CYDV-RPV
T13 Species 533 567 Cucurbit aphid-borne yellows virus
N13 Reference T13 Taxonomy:91753
T14 Species 569 574 CABYV
N14 Reference T14 Taxonomy:91753
T15 Species 577 598 Potato leafroll virus
N15 Reference T15 Taxonomy:12045
T16 Species 600 604 PLRV
N16 Reference T16 Taxonomy:12045
T17 Species 610 630 Turnip yellows virus
N17 Reference T17 Taxonomy:131083
T18 Species 632 636 TuYV
N18 Reference T18 Taxonomy:131083
T19 Species 1141 1149 CYDV-RPV
T20 Species 1154 1158 TuYV
N20 Reference T20 Taxonomy:131083
T21 Species 1222 1226 SbDV
N21 Reference T21 Taxonomy:12049
T22 Species 1228 1232 BLRV
N22 Reference T22 Taxonomy:12041
T23 Species 1234 1238 BChV
N23 Reference T23 Taxonomy:131082
T24 Species 1240 1244 BMYV
N24 Reference T24 Taxonomy:156690
T25 Species 1246 1251 CABYV
N25 Reference T25 Taxonomy:91753
T26 Species 1263 1267 PLRV
N26 Reference T26 Taxonomy:12045
T27 Species 1271 1275 BWYV
N27 Reference T27 Taxonomy:12042
N11 Reference T11 Taxonomy:2170100 Cereal yellow dwarf virus RPV
N12 Reference T12 Taxonomy:2170100 Cereal yellow dwarf virus RPV
N19 Reference T19 Taxonomy:2170100 Cereal yellow dwarf virus RPV