A Rapid and Precise Diagnostic Method for Detecting the Pinewood Nematode <i>Bursaphelenchus xylophilus</i> by Loop-Mediated Isothermal Amplification

Kikuchi, Taisei; Aikawa, Takuya; Oeda, Yuka; Karim, Nurul; Kanzaki, Natsumi

doi:10.1094/phyto-99-12-1365

Cited by 179 publications

(118 citation statements)

References 15 publications

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“…However, it can also be compared to numerous other species-specific molecular detection or identification methods that identify species such as polymerase chain reaction (PCR) (Kiewnick et al 2013), real-time PCR (qPCR) (Huang et al 2010), and isothermal amplification techniques such as loop-mediated isothermal amplification (LAMP) (Kikuchi et al 2009). However, the latter methods are typically targeted to a single species (or occasionally a small group of species), and therefore they are used in the context of answering the question "is this specimen species x?".…”

Section: Introductionmentioning

confidence: 99%

DNA barcoding for biosecurity: case studies from the UK plant protection program

Hodgetts

Ostojá-Starzewski

Prior

et al. 2016

Genome

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Since its conception, DNA barcoding has seen a rapid uptake within the research community. Nevertheless, as with many new scientific tools, progression towards the point of routine deployment within diagnostic laboratories has been slow. In this paper, we discuss the application of DNA barcoding in the Defra plant health diagnostic laboratories, where DNA barcoding is used primarily for the identification of invertebrate pests. We present a series of case studies that demonstrate the successful application of DNA barcoding but also reveal some potential limitations to expanded use. The regulated plant pest, Bursephalenchus xylophilus, and one of its vectors, Monochamus alternatus, were found in dining chairs. Some traded wood products are potentially high risk, allowing the movement of longhorn beetles; Trichoferus campestris, Leptura quadrifasciata, and Trichoferus holosericeus were found in a wooden cutlery tray, a railway sleeper, and a dining chair, respectively. An outbreak of Meloidogyne fallax was identified in Allium ampeloprasum and in three weed species. Reference sequences for UK native psyllids were generated to enable the development of rapid diagnostics to be used for monitoring following the release of Aphalara itadori as a biological control agent for Fallopia japonica.Key words: plant health, regulated quarantine pest, DNA barcoding, diagnostics, invertebrate.Résumé : Depuis son développement, le codage à barres de l'ADN a connu une adoption rapide au sein de la communauté scientifique. Néanmoins, comme pour plusieurs nouveaux outils scientifiques, les avancées en vue d'un usage routinier au sein de laboratoires de diagnostic ont été lentes. Dans ce travail, les auteurs discutent de l'emploi du codage à barres au sein des laboratoires de diagnostique phytosanitaire de DEFRA, où le codage à barres est principalement employé pour l'identification des ravageurs invertébrés. Les auteurs présentent une série d'études de cas le codage à barres a été mis en oeuvre avec succès tout en révélant des limitations potentielles à son usage accru. Le parasite réglementé, Bursephalenchus xylophilus, et l'un de ses vecteurs, Monochamus alternatus, ont été retrouvés dans des chaises de salle à manger. Certains produits du bois sont potentiellement à haut risque en facilitant le mouvement des longicornes; Trichoferus campestris, Leptura quadrifasciata et Trichoferus holosericeus ont été trouvés, respectivement, dans un range-couvert en bois, une traverse ferroviaire, et une chaise de salle à manger. Une épidémie du Meloidogyne fallax a été identifiée chez l'Allium ampeloprasum et chez trois adventices. Des séquences de référence pour des psylles indigènes du Royaume-Uni ont été générées pour permettre le développe-ment d'outils diagnostiques rapides en vue de surveiller l'Aphalara itadori suite à son introduction en tant qu'agent de lutte biologique contre le Fallopia japonica. [Traduit par la Rédaction] Mots-clés : santé des végétaux, organisme nuisible réglementé, codage à barres de l'ADN, diagnostique...

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Section: Introductionmentioning

confidence: 99%

DNA barcoding for biosecurity: case studies from the UK plant protection program

Hodgetts

Ostojá-Starzewski

Prior

et al. 2016

Genome

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“…One previously described method, an adaptation of the cetylmethylammonium bromide (CTAB) method, was efficient, but laborious and time consuming (Takeuchi et al 2005;Futai 2007, 2009). Another method described by Kikuchi et al (2009) and Kanetani et al (2011), cannot be reproduced as there is no description of the buffers and the mentioned kit is not commercially available. The DNA extraction methods, directly from wood samples, described by Wang et al (2010) and Hu et al (2011) are time consuming and use only 5 mg of wood sample as starter material, which is a much smaller sample than the 100 mg used in this study.…”

Section: Discussionmentioning

confidence: 99%

“…Molecular detection methods are simpler, faster and reliable and thus several PCR based methods to detect B. xylophilus with species-specific primers have been developed by targeting either rDNA, ITS regions, satDNA or Hsp70 and DNA topoisomerase I genes (Iwahori et al 1998(Iwahori et al , 2000Kang et al 2004;Matsunaga and Togashi 2004;Cao et al 2005;Castagnone et al 2005;Leal et al 2005Leal et al , 2007Kang et al 2009;Huang et al 2010;Zhuo et al 2011). However, few studies on the use of these methodologies for direct detection of PWN in pine wood and insect vector, without the preliminary steps of nematode extraction, have been conducted (François et al 2007;Takeuchi and Futai 2007;Kikuchi et al 2009;Takeuchi and Futai 2009;Kanetani et al 2011;Wang et al 2010;Hu et al 2011;Wang et al 2011). In the present study, a new methodology was developed for the direct detection of PWN, performed by PCR amplification of the species specific MspI satDNA, leading to a pattern of monomer and multimers of the 160 bp monomer (Tarés et al 1994;Castagnone et al 2005), as recommended by the European and Mediterranean Plant Protection Organization (EPPO) (2009), using total DNA extracted from P. pinaster wood and bark samples and from the insect vector, M. galloprovincialis.…”

Section: Introductionmentioning

confidence: 99%

Direct molecular detection of the pinewood nematode, Bursaphelenchus xylophilus, from pine wood, bark and insect vector

2011

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The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. The international economic impact of the introduction of the PWN into new areas has highlighted the need for the development of accurate and reliable detection methods of B. xylophilus, which are essential to define aspects of its control and management. In the present study, a methodology was developed for the direct detection of PWN by conventional PCR assay, with a species specific set of primers based on PWN satellite DNA, using total DNA extracted directly from maritime pine, Pinus pinaster, wood and bark samples, and from the insect vector, Monochamus galloprovincialis. This methodology involves homogenisation of wood, bark and insects using liquid nitrogen, DNA extraction and one or two PCR amplification steps, which permit the rapid and direct detection of one single nematode present in 100 mg of wood and bark and in one entire insect without the preliminary steps of nematode extraction.

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“…Early on-site detection of plant pathogens with portable biosensors will enable the design of strategies to control the spread of diseases and will also help the study of disease epidemiology. 156 The agricultural sector today requires biosensors for detection of multiple potential pathogens or newly emerged pathogens 157 without necessarily knowing the plant diseases affecting that particular crop group. The development of portable and rapid plant pathogen biosensors would be of value to users from various backgrounds, including individual growers, regulatory agencies, exporters, and researchers.…”

Section: Plant Pathogen Detectionmentioning

confidence: 99%

Development and Applications of Portable Biosensors

2015

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The significance of microfluidics-based and microelectromechanical systems-based biosensors has been widely acknowledged, and many reviews have explored their potential applications in clinical diagnostics, personalized medicine, global health, drug discovery, food safety, and forensics. Because health care costs are increasing, there is an increasing need to remotely monitor the health condition of patients by point-of-care-testing. The demand for biosensors for detection of biological warfare agents has increased, and research is focused on ways of producing small portable devices that would allow fast, accurate, and on-site detection. In the past decade, the demand for rapid and accurate on-site detection of plant disease diagnosis has increased due to emerging pathogens with resistance to pesticides, increased human mobility, and regulations limiting the application of toxic chemicals to prevent spread of diseases. The portability of biosensors for onsite diagnosis is limited due to various issues, including sample preparation techniques, fluid-handling techniques, the limited lifetime of biological reagents, device packaging, integrating electronics for data collection/analysis, and the requirement of external accessories and power. Many microfluidic, electronic, and biological design strategies, such as handling liquids in biosensors without pumps/valves, the application of droplet-based microfluidics, paper-based microfluidic devices, and wireless networking capabilities for data transmission, are being explored.

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A Rapid and Precise Diagnostic Method for Detecting the Pinewood Nematode Bursaphelenchus xylophilus by Loop-Mediated Isothermal Amplification

Cited by 179 publications

References 15 publications

DNA barcoding for biosecurity: case studies from the UK plant protection program

DNA barcoding for biosecurity: case studies from the UK plant protection program

Direct molecular detection of the pinewood nematode, Bursaphelenchus xylophilus, from pine wood, bark and insect vector

Development and Applications of Portable Biosensors

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