A Rapid and Precise Diagnostic Method for Detecting the Pinewood Nematode Bursaphelenchus xylophilus by Loop-Mediated Isothermal Amplification
A Rapid and Precise Diagnostic Method for Detecting the Pinewood Nematode Bursaphelenchus xylophilus by Loop-Mediated Isothermal Amplification
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DOI：
10.1094/phyto-99-12-1365
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发表时间：
2009-12-01
期刊：
影响因子：
3.2
通讯作者：
Kanzaki, Natsumi
中科院分区：
文献类型：
--
作者：
Kikuchi, Taisei;Aikawa, Takuya;Kanzaki, Natsumi
松材线虫病是日本、韩国、中国、台湾和葡萄牙的主要森林病害。一种快速、准确、简单的诊断方法可以极大地帮助这种疾病的正确管理。在这里，我们提出了一种新的检测方法，使用环介导的等温扩增（LAMP）靶向的核糖体DNA的内部转录间隔区的线虫。使用来自与B相关的各种线虫物种的DNA证实引物和LAMP的特异性。木生植物我们的实验结果表明，LAMP可以检测到B。与传统的诊断方法相比，具有更快、更高的灵敏度。此外，由于它不需要昂贵的设备或专门的技术，这种基于LAMP的诊断方法具有在现场条件下使用的潜力。
Bursaphelenchus xylophilus is the causal agent of pine wilt disease, which is a major forest disease in Japan, Korea, China, Taiwan, and Portugal. A diagnostic method which is rapid, precise, and simple could greatly help the proper management of this disease. Here, we present a novel detection method using loop-mediated isothermal amplification (LAMP) targeting the internal transcribed spacer region of ribosomal DNA of the nematode. Specificity of the primers and LAMP was confirmed using DNA from various nematode species related to B. xylophilus. Our experimental results suggest that LAMP can detect B. xylophilus faster and with higher sensitivity than the traditional diagnostic method. Moreover, because it does not require expensive equipment or specialized techniques, this LAMP-based diagnostic method has the potential to be used under field conditions.