数字农科院2.0

A one-step multiplex RT-PCR assay for simultaneous detection of four viruses that infect peach

文献类型： 外文期刊

作者： Zhao, Z.;Jiang, D.;Li, S.;Yu, Y.;Jiang, D.;Wu, Z.;Zhao, Z.

作者机构：

关键词： detection assays;ELISA;multiplex RT-PCR;peach viruses

期刊名称： LETTERS IN APPLIED MICROBIOLOGY

ISSN： 0266-8254

年卷期： 2013 年 57 卷 4 期

页码：

收录情况： SCI

摘要： A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed to enable the simultaneous detection and differentiation of four viruses that infect peach, namely Apple chlorotic leaf spot virus (ACLSV), Cherry green ring mottle virus (CGRMV), Prunus necrotic ringspot virus (PNRSV) and Apricot pseudo-chlorotic leaf spot virus (APCLSV). In this study, four pairs of primers, one specific for each virus, were designed; the corresponding PCR products were 632, 439, 346 and 282bp in length for ACLSV, CGRMV, PNRSV and APCLSV, respectively, and the fragments could be distinguished clearly by agarose gel electrophoresis. The sensitivity and specificity of the method were tested using individual RT-PCR and enzyme-linked immunosorbent assay (ELISA), and the identity of the RT-PCR amplification products was also confirmed by DNA sequencing. The results of RT-PCR and ELISA, along with batch detection using samples collected from peach orchards, revealed that this rapid and simple technique is an effective way to identify the four viruses simultaneously.

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