关键词: 美国白蛾核型多角体病毒；SYBR Green &Iota, ；荧光定量PCR；感病幼虫；检测

Abstract: Primers were designed based on polyhedrin from Hyphantria cunea nucleopolyhedrovirus in order to establish the detection method of SYBR GreenⅠ real-time fluorescence quantitative PCR for HcNPV, and the standard curve of SYBR GreenⅠreal-time fluorescence quantitative PCR for polyhedrin was established through PCR amplification, ligation of target genes and vector, transformation, identification of recombinant plasmid and DNA sample detection. Statistic analysis showed that there was a good linear relationship between Ct value and the logarithmic value of plasmid concentrations （R=0.998）. The sensitivity of the method was 10¹-1×10² copies/μL, and wide detection range of 5 orders of magnitude was obtained. Larvae infected at different time were sampled and detected by the method, and the results showed that there was a good linear relation between the logarithmic value of the multiples of gene copies and the infection time （R= 0.987）. The results show that the detection method for HcNPV is of high sensitivity, reproducibility and credibility, which will facilitate the epizootiology and the standardized production for insect virus.  

Key words: Hyphantria cunea nucleopolyhedrovirus, SYBR Green I, fluorescence quantitative PCR, infected larvae, detection