Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan®MGB probes

被引：27
|
作者：
Debreczeni, D. E. [1 ]
Ruiz-Ruiz, S. [1 ]
Aramburu, J. [2 ]
Lopez, C. [3 ]
Belliure, B. [1 ]
Galipienso, L. [2 ]
Soler, S. [3 ]
Rubio, L. [1 ]
机构：
[1] IVIA, Valencia 46113, Spain
[2] IRTA, Barcelona 08348, Spain
[3] Univ Politecn Valencia, COMAV, Ctr Conservac & Mejora Agrodiversidad Valenciana, Valencia 46022, Spain
来源：
JOURNAL OF VIROLOGICAL METHODS | 2011年 / 176卷 / 1-2期
关键词：
TSWV; Tospovirus; Diagnosis; Quantitative RT-PCR; CITRUS-TRISTEZA-VIRUS; FRANKLINIELLA-OCCIDENTALIS; EVOLUTIONARY ANALYSIS; GENETIC-VARIATION; ASSAY; RESISTANCE; TSWV; EPIDEMICS; FITNESS; PROTEIN;
D O I：
10.1016/j.jviromet.2011.05.027
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) procedure using a general primer set and three TaqMan (R) MGB probes was developed for general and genotype-specific detection and quantitation of the genomic M segment of Tomato spotted wilt virus (TSWV). Standard curves using RNA transcripts homologous to the three probes allowed reproducible quantitative assays with a wide dynamic range (10(3)-10(10) TSWV M segment RNA copies/ng of total RNA) and high sensitivity. This protocol was assayed with a battery of TSWV isolates, covering the range of the present known genetic variation, in single and/or mix infections in three plant hosts, as well as in the thrips vector Frankliniella occidenialis. This quantitative detection assay will be a valuable tool for molecular biology and epidemiology studies, diagnosis and disease control. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码：32 / 37
页数：6
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