Multiplex RT-PCR detection of four aphid-borne strawberry viruses in Fragaria spp. in combination with a plant mRNA specific internal control
Multiplex RT-PCR detection of four aphid-borne strawberry viruses in Fragaria spp. in combination with a plant mRNA specific internal control
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DOI：
10.1016/s0166-0934(03)00164-2
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发表时间：
2003-08-01
影响因子：
3.1
通讯作者：
Jelkmann, W
中科院分区：
文献类型：
--
作者：
Thompson, JR;Wetzel, S;Jelkmann, W
感染草莓（Fragaria spp.）的主要蚜虫传播病毒有草莓皱纹病毒（SCV）、草莓淡黄边缘病毒（SMYEV）、草莓斑驳病毒（SMoV）和草莓脉带病毒（SVBV），可造成严重的农作物损失。本文描述了一种多重逆转录酶聚合酶链式反应 (RT-PCR) 方法，用于同时检测所有四种病毒，并结合植物 mRNA 特异性内部对照，该内部对照可用作提取和 RT-PCR 有效性的指标。通过该方法总共分析了 18 个自然感染的草莓分离株。 RNA 病毒的每种组合都能够被检测到，并且在三个分离株中发现了所有四种病毒的完整互补体，所有分离株均取自智利的野草莓（Fragaria chiloensis (L.) Duch.）。四种病毒的检测上限为提取物稀释度为 1/200。 RNA 特异性内部对照引物的广泛适用性（在测试的 27 种植物中的 25 种中产生了预期大小的 PCR 片段）与所述提取方法中的改进相结合，为多种植物物种中可比较的 RT-PCR 分析提供了潜在的标准方法。 (C) 2003 Elsevier B.V. 保留所有权利。
The principal aphid-borne viruses infecting Strawberry (Fragaria spp.) Strawberry crinkle virus (SCV), Strawberry mild yellow, edge virus (SMYEV), Strawberry mottle virus (SMoV) and Strawberry vein banding virus (SVBV) can cause serious crop losses. In this paper, a multiplex reverse transcriptase polymerase chain reaction (RT-PCR) method is described for the simultaneous detection of all four viruses in combination with a plant mRNA specific internal control which can be used as an indicator of the effectiveness of the extraction and RT-PCR. In total, 18 strawberry isolates infected naturally were analysed by this method. Every combination of RNA virus was able to be detected and a full complement of all four viruses were found together in three isolates, all taken from wild strawberry (Fragaria chiloensis (L.) Duch.) in Chile. The upper detection limit for the four viruses was at an extract dilution of 1/200. The broad applicability of the RNA specific internal control primers-which produced a PCR fragment of the expected size in 25 of 27 plant species tested-combined with improvements, made in extraction methods described provides potentially a standard method for comparable RT-PCR analyses in a wide variety of plant species. (C) 2003 Elsevier B.V. All rights reserved.