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首页> 外文期刊>PLoS One >Point-of-care diagnostic (POCD) method for detecting Bursaphelenchus xylophilus in pinewood using recombinase polymerase amplification (RPA) with the portable optical isothermal device (POID)
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Point-of-care diagnostic (POCD) method for detecting Bursaphelenchus xylophilus in pinewood using recombinase polymerase amplification (RPA) with the portable optical isothermal device (POID)

机译：用便携式光学等温装置（POID）使用重组酶聚合酶扩增（RPA）在Pinewood中检测松下的诊断（POCD）方法

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摘要

The pinewood nematode (PWN), Bursaphelenchus xylophilus , is a causative agent of pine wilt disease (PWD). To date, although several molecular diagnostic methods have been developed, rapid on-site diagnostic tools for detecting PWN in pinewood are limited. In this study, a point of care diagnostic (POCD) method for detecting PWN in pinewood using recombinase polymerase amplification (RPA) assay was developed. This method comprises quick gDNA extraction buffer (DAP buffer) for the direct extraction of gDNA of PWN from pinewood and a battery-mounted portable optical isothermal device (POID) for the detection of PWD in the field. The RPA assay can distinguish between the PWN and its conspecies which exist in pinewood and can complete diagnostic procedures within 25 min in the field. Moreover, the RPA assay can detect PWN in old wood samples in both natural and stored conditions. The POCD-RPA assay to detect PWN will be useful for epidemiological investigations in the field as well as for quarantine processes in the wood trade.
机译：Pinewood Nematode（PWN），Bursaphelenchus Xylophilus是松树枯萎病（PWD）的致病因子。迄今为止，虽然已经开发了几种分子诊断方法，但是有限地检测PWN的快速现场诊断工具。在本研究中，开发了使用重组酶聚合酶扩增（RPA）测定的用于检测Pinewood中PWN的护理诊断（POCD）方法。该方法包括快速GDNA提取缓冲液（DAP缓冲液），用于从松木和电池安装的便携式光学等温装置（POID）直接提取PWN的GDNA，用于检测该领域的PWD。 RPA测定可以区分PINEWOOD中存在的PWN及其终结，并且可以在现场25分钟内完成诊断程序。此外，RPA测定可以在天然和储存条件下检测旧木样品中的PWN。 POCD-RPA测定检测PWN将可用于该领域的流行病学调查以及木材贸易中的检疫过程。

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