Welcome to visit Scientia Silvae Sinicae，Today is

Scientia Silvae Sinicae ›› 2005, Vol. 41 ›› Issue (2): 91-95.doi: 10.11707/j.1001-7488.20050215

Previous Articles     Next Articles

Development and Application of an Immunoassay Method for Identification of Bursaphelenchus xylophilus in Pinewood

Bai Gang1,Wang Yuyan2,Ma Hongzhou1,Li Haiyan2,Yang Wenbo1   

  1. 1.College of Life Science, Nankai University Tianjin 300071; 2.General Station of Forest Pest and Disease Control, The State Administration of Forestry Shenyang 11003
  • Received:2003-03-03 Revised:1900-01-01 Online:2005-03-25 Published:2005-03-25

PDF (PC)

675

Abstract:

Taking the specimens of Bursaphelenchus xylophilus (BX) and B. mucronatus (BM) fragments as research objects, an immunoassay method for nematode antigen analysis was developed in this paper. The rabbit antiserum against BX was prepared, and the antigen proteins were assayed by SDS-PAGE and western blot. Using BX fragments as solid-phase, the assay principle was based on competition between an analyte and nematode solid-phase antigen for anti-BX antibody. Then using HRP labeled goat anti-rabbit IgG as a tracer antibody, an enzyme-linked immunosorbent assay (ELISA) was developed. The method was specific to BX antigen harvested from different areas. Although the ELISA showed a little cross reactivity with BM, it could identify 0.1μg nematode protein in 10 mg pinewood specimen. The result suggested that the ELISA method using nematode antigen as a detection objective showed a new potential for the quarantine inspection and forest pest control of BX.

Key words: Bursaphelenchus xylophilus, nematode analysis, western blot, ELISA