湖北林业科技

In this paper, a new two gene-jointed PCR detection assay to detect Bursaphelenchus xylophilus, was estab- lished. For developing the two gene-jointed PCR detection of B. xylophilus, two pairs of primers were designed according to sequence of B. xylophilus available in GenBank, targeting the cathepsin L-like cysteine proteinase gene and ribosomal DNA internal transcribed spacer region . The specificity, sensitivity assay and clinical samples were tested by using the opti- mized reaction system . Results showed that the specific fragments 490bp and 264 bp were able to amplified, while there was no amplification product found in positive controls and other tested ones . The method was applied to detect clinical samples and the result showed 100% consistence with PCR. These results could be served as a basis detection application in diagnosis of B. xylophilus.