PCR-based Rapid Detection Of Two Baculoviruses Of Forest Insects

Forest insect virus as a highly efficient sound-specific insecticide, is played a significant role in the forest protection now. But it is not be used popularly, mainly because virus has a long latent period, its reaction are slower than chemical pesticides, and these questioned the users. This experiment aimed at establishing several insect virus nucleic acid-based detection technologies, which fill the blank of the insect virus detection in the molecular biology. Applied the technologies to detect low concentration of OB suspension can provide some evidence for vertical transmission and reaction effect of insect virus, it also can demonstrate the feasibility of controlling pest with insect virus.Two pairs of specific primers were used for the detection of the Hyphantria cunea nucleopolyhedrovirus (HcNPV) by PCR. The two pairs of primers were both designed based on gene pe38 of HcNPV genome. The sizes of amplified fragments were 994 and 614 bp, respectively. The two pairs of primers were further used to detect the amount of viral DNA in diseased larvae or polyhedron of HcNPV. The minimum amount of detection could be as less as 1 fg of total DNA or 3?4 OBs/mL of polyhedron. The amount of viral DNA in hemolymph sampled from the larvae infected by different concentrations of polyhedron at different time points was also investigated, and it was found that the lower the concentration of polyhedron used, the later the viral DNA will be detected.Larvae of Hyphantria cunea were collected from Qinhuangdao. Their hemolymph is detected by PCR, the effect is also obviously. The result shows that the PCR-based method can be applied to practice, and HcNPV has a continuous control to Hyphantria cunea.Two pairs of specific primers were used for the detection of the Clostera anachoreta Granulovirus (ClanGV) by PCR. The two pairs of primers were both designed based on gene egt of ClanGV genome. The sizes of amplified fragments were 401 and 491 bp, respectively. The two pairs of primers were used to detect the genome DNA and OBs of ClanGV successfully.