›› 2009, Vol. 52 ›› Issue (6): 707-712.

• 研究论文 • 上一篇    


王文欢, 曲良建, 王玉珠, 张永安   

  • 出版日期:2009-06-20 发布日期:2009-06-20
  • 通讯作者: 张永安

PCR-based early detection of Hyphantria cunea nucleopolyhedrovirus (HcNPV)

  • Online:2009-06-20 Published:2009-06-20

摘要: 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614 bp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1 fg病虫总DNA和3~4 OBs/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3.53×109 OBs/mL时, 24 h后可检出病毒DNA;接毒量为3~4 OBs/mL时, 120 h后可检出。病毒可检出时间随接毒浓度的递减而延后。

关键词: 美国白蛾核型多角体病毒, 生物防治, 早期检测, PCR, pe38基因

Abstract: Two pairs of specific primers were used for the detection of 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 low 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.

Key words: Hyphantria cunea nucleopolyhedrovirus (HcNPV), biocontrol, early detection, PCR, pe38 gene