Pine wilt disease (PWD) induced by the pinewood nematode (PWN), Bursaphelenchus xylophilus, is a great threat to pine forests in Europe and East Asia. Identification of interrelation between Monochamus spp., the major vectors of PWD and PWN is the key factor for effective control of PWD because PWN is trasmitted to healthy pine trees only by Monochamus vectors. In this study, direct injection system is designed to apply PWN artificially to M. alternatus just before emergence to adults. After injection of PWN (300~500 nematodes in 10 μl of 1X PBS) using adapted glass pipets, the survival rate of M. alternatus and detection rate of live PWN were 80.0% and 68.8% at 168 h respectively. As a result, we demonstrated the artificial injection methods of nematode to Monochamus vectors for further study of coeffect between vectors and specific stage of PWN including 3rd stage dispersal juvenile.