Molecular diagnosis of Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle (Nematoda Aphelenchida Parasitaphelenchidae) in presence of polyphenols and other inhibitory compounds.

Abstract

The pine wood nematode Bursaphelenchus xylophilus (Aphelenchida Parasitaphelenchidae) infects conifers, in particular pines, causing pine wilt disease. This nematode is considered a quarantine organism according to the EPPO and AQISQ guidelines. For this reason, several procedures for an affordable diagnostic test have been developed based on LAMP technique. In this work we shown how this technique behaved in presence of compounds that exert an inhibitory effect on DNA polymerases, a common problem that could be found in DNA preparations obtained from samples included in plant matrixes  LAMP technique was found able to detect presence of its specific DNA target in almost all cases studied but we found that the addiction of polyphenolic compounds present in some woody matrixes to DNA preparations could affect significantly on the sensibility of diagnostic test, while resinous material had a lesser influence on amplification.