Application of Multiplex TaqMan Real-Time PCR Assay in Survey of Five Lily Viruses Infecting Lilium spp.

文献类型： 外文期刊

第一作者： Xu, Leifeng

作者： Xu, Leifeng;Song, Meng;Ming, Jun

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关键词： Lilium spp; LSV; LMoV; CMV; SYSV; PlAMV; multiplex TaqMan real-time PCR; simultaneous detection

期刊名称：AGRONOMY-BASEL （ 影响因子：3.949； 五年影响因子：4.117 ）

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年卷期： 2022 年 12 卷 1 期

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收录情况： SCI

摘要： Lily symptomless virus (LSV), Lily mottle virus (LMoV), Cucumber mosaic virus (CMV), Shallot yellow stripe virus (SYSV), and Plantago asiatica mosaic virus (PlAMV) are five of the economically important viruses infecting lilies (Lilium spp.) worldwide. In order to prevent the occurrence and spread of these viruses, it is necessary to develop a rapid, effective, and sensitive detection method for the simultaneous detection and specific quantification of these viruses. In this study, specific primers and probes for multiplex TaqMan real-time PCR assays designed from conserved regions of the coat protein sequence of each virus were used for the simultaneous detection of these viruses in lilies (Lilium spp.). The optimal concentration of primers and probes and reaction annealing temperature were 20 mu M and 55.9 degrees C, respectively. The detection limits of the assay were 1.33 x 10(2), 1.27 x 10(1), 1.28 x 10(1), 2.33 x 10(2), and 2.01 x 10(2) copies center dot mu L-1 for LSV, LMoV, CMV, SYSV, and PlAMV, respectively. Specificity was determined using seven viral pathogens of lilies. Variability tests of intra- and inter-assays showed high reproducibility with coefficients of variation <2%. The multiplex TaqMan real-time PCR assay was used to detect these viruses from lily samples in China. In brief, our developed assay showed high specificity, sensitivity, and reproducibility for the simultaneous detection and differentiation of five lily-infecting viruses and can be used for certification and quarantine programs.

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