Detection of four regulated grapevine viruses in a qualitative, single tube real-time PCR with melting curve analysis

被引：10

作者：

Aloisio, M. ^{[1
,2
]}

Morelli, M. ^{[3
]}

Elicio, V. ^{[4
]}

Saldarelli, P. ^{[3
]}

Ura, B. ^{[2
]}

Bortot, B. ^{[2
]}

Severini, G. M. ^{[2
]}

Minafra, A. ^{[3
]}

机构：

[1] Univ Trieste, Dept Life Sci, Trieste, Italy

[2] IRCCS Burlo Garofolo, Inst Maternal & Child Hlth, Trieste, Italy

[3] Inst Sustainable Plant Protect, Bari, Italy

[4] Agritest Srl, Valenzano, Italy

来源：

JOURNAL OF VIROLOGICAL METHODS | 2018年 / 257卷

关键词：

Multiplex Real-time PCR; Melting curve; EvaGreen; Grapevine; Viruses; POLYMERASE CHAIN-REACTION; MULTIPLEX RT-PCR; SYBR GREEN; RAPID DETECTION; ASSAY; DIFFERENTIATION; VALIDATION; EVAGREEN; TYPE-2;

D O I：

10.1016/j.jviromet.2018.04.008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The detection of the four grapevine viruses (GLRaV-1, GLRaV-3, GFLV and ArMV) regulated in European Union plant material certification, requires sensitive and specific diagnostic tools. A strategy of simultaneous detection in a real-time single tube amplification was developed, based on the EvaGreen binding dye. The melting curve analysis (MCA) of the amplicons allows a qualitative detection of the four different virus targets in multiplex analysis. A plasmid dilution assay calculated an analytical sensitivity with an amplification threshold up to 100 copies of the target sequences. A small cohort of field grapevine samples, with a known status of infection by mixtures of the target viruses or free of them, respectively, was successfully tested for the evaluation of the amplicons Tm.

引用

页码：42 / 47

页数：6

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