The use of real-time polymerase chain reaction with high resolution melting (real-time PCR-HRM) analysis for the detection and discrimination of nematodes Bursaphelenchus xylophilus and Bursaphelenchus mucronatus

• The real-time PCR-HRM analysis was developed for the detection and discrimination of Bursaphelenchus xylophilus and Bursaphelenchus mucronatus.
• The results showed distinct melting curve profiles related to each of examined species.
• The method may be used to detect and discriminate these both species, regardless of the life stages of nematodes and template concentrations.

The real-time PCR-HRM analysis was developed for the detection and discrimination of the quarantine nematode Bursaphelenchus xylophilus and Bursaphelenchus mucronatus. A set of primers was designed to target the ITS region of rDNA. The results have demonstrated that this analysis is a valuable tool for differentiation of these both species.