PCR diagnostic method for Bursaphelenchus xylophilus
A simple PCR-RFLP method was developed in Japan to identify Bursaphelenchus xylophilus (EPPO A1 quarantine pest) and differentiate it from B. mucronatus. This method can be used on a single nematode, living or preserved (except nematodes preserved in fixatives containing aldehyde). An individual nematode (juvenile, adult) is crushed with a filter paper chip. This filter paper chip is placed into PCR buffer as the DNA template. The primer set used has been selected to amplify the internal transcribed spacer 1 and 2 regions of 5.8S rDNA. RFLP is then used to differentiate B. xylophilus from B. mucronatus. The authors concluded that their method is simple and reliable.
Iwahori, H.; Kanzaki, N.; Futai, K. (2000) A simple, polymerase chain reaction-restriction fragment length polymorphism-aided diagnosis method for pine wilt disease.
Forest Pathology, 30(3), 157-164.