Point-of-care diagnostic (POCD) method for detecting Bursaphelenchus xylophilus in pinewood using recombinase polymerase amplification (RPA) with the portable optical isothermal device (POID)


Autoři: Deokjea Cha aff001;  Dongsoo Kim aff001;  Wonil Choi aff002;  Sungjun Park aff003;  Hyerim Han aff001
Působiště autorů: Division of Forest Insect Pests & Diseases, National Institute of Forest Science, Dongdaemun, Seoul, Korea aff001;  Division of Forest Ecology & Climate Change, National Institute of Forest Science, Dongdaemun, Seoul, Korea aff002;  2NCBio, Seo-gu, Daejeon, Korea aff003
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0227476

Souhrn

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.

Klíčová slova:

Forests – Nematode infections – Optical equipment – Pines – Polymerase chain reaction – Recombinase polymerase amplification – Sequence alignment – Trees


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2020 Číslo 1