HORSERADISH PEROXIDASE-LABELED PROBES AND ENHANCED CHEMILUMINESCENCE TO DETECT BACULOVIRUSES IN GYPSY-MOTH AND EASTERN SPRUCE BUDWORM LARVAE

Horseradish peroxidase-labelled whole genomic DNA probes and enhanced chemiluminescence procedures were utilized to detect baculovirus in in sect macerates blotted on nylon membranes. Detection levels were simil ar to those found using P-32-labelled probes; 5 . 10(3) occlusion bodi es (OBs), 2. 10(3) OBs and 4 . 10(4) OBs of Lymantria dispar (L.) nucl ear polyhedrosis virus (LdNPV), Choristoneura fumiferana (Clem.) NPV ( CfNPV) and C. fumiferana granulosis virus (CfGV) respectively using 10 ng/ml LdNPV DNA probe and 20 ng/ml CfNPV and CfGV probe concentration s. Quantities of purified viral DNA detected were 0.56 ng LdNPV, 0.20 ng CfNPV and 0.10 ng CfGV at similar probe concentrations. Cross react ions were observed between LdNPV DNA probes and CfNPV. Multiple probin g of membranes blotted with insect macerates was capable of diagnosing the presence of NPV and GV on membranes. This procedure appears to be useful in the diagnosis of large numbers of insects for several bacul oviruses.