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用于检测马铃薯黄脉病毒(PYVV)的实时和常规逆转录聚合酶链反应检测方法的开发。

Development of real-time and conventional RT-PCR assays for the detection of potato yellow vein virus (PYVV).

作者信息

López R, Asensio C, Guzman M M, Boonham N

机构信息

Central Science Laboratory, Sand Hutton, York YO41 1LZ, UK.

出版信息

J Virol Methods. 2006 Sep;136(1-2):24-9. doi: 10.1016/j.jviromet.2006.03.026. Epub 2006 May 18.

Abstract

Potato yellow vein virus (PYVV) is considered a quarantine pathogen in the European and Mediterranean Plant Protection Organization (EPPO) area. This virus is widespread and damaging at its centre of origin in South America. Current detection methods are either time-consuming or difficult to interpret. This paper reports the development of a sensitive, high throughput, real-time reverse transcription (RT)-PCR assay, based on TaqMan chemistry, suitable for PYVV detection. In addition, a reliable conventional RT-PCR assay for PYVV detection is also presented. Although less sensitive (1000 times less sensitive in direct comparison), this method requires less sophisticated equipment and as such should be a useful alternative to the real-time technique in some testing laboratories. The two assays presented here could assist in the implementation of quarantine measures for PYVV identification and in routine indexing of PYVV for the production of virus-free seed potatoes in areas of South America where the virus is highly damaging.

摘要

马铃薯黄脉病毒(PYVV)在欧洲和地中海植物保护组织(EPPO)区域被视为检疫性病原菌。该病毒在其南美洲的起源中心广泛传播且具有危害性。目前的检测方法要么耗时,要么难以解读。本文报道了一种基于TaqMan化学的灵敏、高通量实时逆转录(RT)-PCR检测方法的开发,适用于PYVV检测。此外,还介绍了一种用于PYVV检测的可靠常规RT-PCR检测方法。尽管该方法灵敏度较低(直接比较时低1000倍),但所需设备不那么复杂,因此在一些检测实验室中应是实时技术的有用替代方法。这里介绍的两种检测方法有助于实施针对PYVV鉴定的检疫措施,以及在南美洲该病毒危害严重地区对用于生产无病毒种薯的PYVV进行常规检测。

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