Hassan Mohamed, Myrta Arben, Polak Jaroslav
Research Institute of Crop Production, Department of Virology, Prague, Czech Republic.
J Virol Methods. 2006 May;133(2):124-9. doi: 10.1016/j.jviromet.2005.11.002. Epub 2005 Dec 5.
A pentaplex reverse-transcription polymerase chain reaction (Pentaplex RT-PCR) in a single tube was developed for the simultaneous detection of the pome fruit viruses: Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV) and Apple mosaic virus (ApMV). This is the first report of the simultaneous detection of all four viruses and host mRNA as an internal specific control. Pentaplex RT-PCR was applied successfully throughout the year, using different plant organs (leaves or dormant buds). The sensitivity of detection by monoplex- and pentaplex RT-PCR assays was comparable. Different combinations of mixed infections of viruses were identified in samples of infected apple and pear trees from different geographical regions. The pentaplex RT-PCR assay developed was sensitive, simple, rapid, and reliable for simultaneous detection of the four viruses in extracts of leaves or dormant buds.
开发了一种单管五重逆转录聚合酶链反应（Pentaplex RT-PCR），用于同时检测核果类果树病毒：苹果茎痘病毒（ASPV）、苹果茎沟病毒（ASGV）、苹果褪绿叶斑病毒（ACLSV）和苹果花叶病毒（ApMV）。这是首次同时检测所有四种病毒以及宿主mRNA作为内部特异性对照的报告。全年使用不同的植物器官（叶片或休眠芽）成功应用了五重RT-PCR。单重和五重RT-PCR检测的灵敏度相当。在来自不同地理区域的感染苹果树和梨树样本中鉴定出了病毒混合感染的不同组合。所开发的五重RT-PCR检测方法灵敏、简单、快速且可靠，可用于同时检测叶片或休眠芽提取物中的四种病毒。
