Detection of living Bursaphelenchus xylophilus in wood, using reverse transcriptase loop‐mediated isothermal amplification (RT‐LAMP)

• DOI: 10.1111/efp.12149
• Publication Date: 2014-10-08T06:51:32Z
• Authors: I. Leal; E. Allen; B. Foord; J. Anema; C. Reisle; A. Uzunovic; A. Varga; D. James
• Keywords: Wilt disease; Phytosanitary certification

Summary Pinewood nematode (PWN), Bursaphelenchus xylophilus , the causal agent of pine wilt disease, is an inhabitant native species North America, where its presence has minor impact. In contrast, introduction this to forests in Asia and Europe devastated some stands recognized as a pest significant phytosanitary concern by National Plant Protection Organizations several countries. The ability detect PWN internationally traded wood products crucial reduce spread organism. Currently, majority molecular techniques for detection rely on genomic DNA thus fail differentiate between living dead PWN. nematodes could lead unnecessary trade disruption. Therefore, accurate differentiation are critical. We have developed reverse transcription loop‐mediated isothermal amplification (RT‐LAMP) assay, which specifically identifies detecting mRNA encoding expansin gene viability marker. This diagnostic method was found be more sensitive, faster less dependent expensive laboratory equipment than PCR. addition, unlike PCR, it allows simple colour products. will help resolve disputes over clarifying whether originates from live or organisms. Where approved treatments implemented, disruption avoided, protecting market access PWN‐infested areas.