Pine wilt disease (PWD), caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus, leads
to serious losses to pine forestry around the world. Pinus massoniana, which is vulnerable to be
attacked by the PWN, is the dominant species used in pine forestry in China. The objective of this
study is to develop a direct PCR-based method for detecting B. xylophilus in the wood of
P. massoniana without a separate nematode extraction step. A simple procedure was first developed
for isolating B. xylophilus DNA in 5 mg pine wood tissue samples harbouring PWN for detection by
PCR amplification. A B. xylophilus-specific amplicon of 403 bp (DQ855275) was generated by PCR
from the infested wood tissue. The entire procedure can be completed within 5 h with one pair of
primers. This assay can serve as a rapid, cheap and environmentally friendly method to detect
B. xylophilus in samples of P. massoniana
