Latent fruit tree viruses present economic threat to the industry and nurseries as diseases they cause not only reduce fruit quality and production yield, but can also be spread inadvertently through propagation due to the lack of viral symptoms on an infected mother plant. As a result, these viruses require appropriate detection tools for effective management. In this study we developed RT-qPCR assays for the detection of three latent viruses of pome, apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), and apple mosaic virus (ApMV), using the alignment of representative sequences from the NCBI database. The optimized assays were shown to be specific by successfully amplifying the target from positive controls without showing any detectable amplification in negative and non-target controls, and revealed high sensitivity by reliably detecting as low as 101 copies per reaction. The results also demonstrated that both the choice of extraction method and the reagents used for RT-qPCRcould play a critical role in virus detection outcome. These assays were both reliable and robust compared to the extant RT-PCR methods, and they could be a viable tool for making informed management decisions.

中文翻译：

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用于检测三种潜伏病毒的RT-qPCR检测方法的开发。

潜在的果树病毒对工业和苗圃构成经济威胁，因为它们引起的疾病不仅降低了水果质量和产量，而且由于在受感染的母株上缺乏病毒症状，还可能通过繁殖无意传播。结果，这些病毒需要适当的检测工具来进行有效管理。在这项研究中，我们开发了RT-qPCR分析方法，使用代表性序列的比对来检测三种潜伏的波普病毒，苹果绿斑病斑点病毒（ACLSV），苹果茎点病毒（ASPV）和苹果花叶病毒（ApMV）从NCBI数据库。通过从阳性对照成功扩增靶标而未显示阴性对照和非靶标对照中可检测到的扩增，优化的检测方法具有特异性。并通过可靠地检测每个反应低至101份拷贝显示出高灵敏度。结果还表明，提取方法的选择和用于RT-qPCR的试剂都可能在病毒检测结果中发挥关键作用。与现有的RT-PCR方法相比，这些测定既可靠又可靠，并且它们可能是做出明智管理决策的可行工具。