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

页码：32 / 37

页数：6

共 24 条

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