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根据美国白蛾核型多角体病毒(Hyphantria cunea nucleopolyhedrovirus,HcNPV)pe38基因设计两对引物,利用PCR法检测HcNPV基因组DNA,分别扩增出长为994和614bp的片段,经测序确定为HcNPV pe38基因片段。应用两对引物分别检测了美国白蛾病虫的总DNA和获得的病毒多角体,其检测最低量分别为1fg病虫总DNA和3～4OBs/mL。用不同浓度的病毒感染试虫后,不同时间取其血淋巴用PCR方法检测。结果表明,接毒量为3.53×109OBs/mL时,24h后可检出病毒DNA;接毒量为3～4OBs/mL时,120h后可检出。病毒可检出时间随接毒浓度的递减而延后。
Two pairs of primers were designed according to pe38 gene of Hypntria cunea nucleopolyhedrovirus (HcNPV). The genomic DNA of HcNPV was amplified by PCR, and the fragments of 994 bp and 614 bp were amplified respectively and sequenced as HcNPV pe38 gene Fragment. Two pairs of primers were used to detect the total DNA and the virus polyhedra of C. medinalis respectively. The detection limits were 1 fg total DNA and 3 ~ 4 OBs / mL, respectively. Infected with different concentrations of test insects worms at different times to take their hemolymph detected by PCR method. The results showed that the virus DNA could be detected after 24 hours when the virus was 3.53 × 109OBs / mL, and it could be detected after 120 hours when it was 3 ~ 4OBs / mL. The detectable time of the virus is delayed with the decrease of the virus concentration.