Development and Application of Immunomagnetic Separation ELISA for Identification of Bursaphelenchus xylophilus in Pinewood

Using dispersion polymerization,polystyrene magnetic micropsheres as carriers were prepared.By coating with rabbit anti-Bursaphelenchus xylophilus(BX) IgG,the immunomagnetic microspheres(IMMS) could specifically capture nematode antigen on pinewood samples.Taking biotinylated Anti-BX rabbit IgG as a tracer antibody,together with horseradish peroxidase labeled avidin,an enzyme-linked immuosorbent assay(ELISA) on IMMS was developed.Three kinds of pine trees harvested from different areas were detected by this method.Although there were a few crossed reactivity between Bursaphelenchus mucronatus antigen with Anti-BX IgG,the relevance ratio of BX on infected pine trees was 94.1% and the detection limit could be 0.1 μg·mL-1 nematode antigen.The result suggested that immunomagnetic separation enzyme-linked immuosorbent assay(IMS-ELISA) could directly capture trace nematode antigen on pinewood homogenizations.The method showed practicability of the quick quarantine inspection for its speediness,convenience and accuracy.