›› 2005, Vol. 48 ›› Issue (6): 871-875.

• RESEARCH PAPERS • Previous Articles     Next Articles

Detection of proliferation of Bombyx mori nucleopolyhedrovirus in its host by fluorescence quantitative PCR

YAO Qin, GAO Lu, CHEN Ke-Ping, HU Zhi-Gang   

  1. Institute of Life Sciences, Jiangsu University
  • Online:2005-12-29 Published:2005-12-20

Abstract:

To investigate the proliferation of Bombyx mori nucleopolyhedrovirus (BmNPV) in its host, the 5th instar larvae of the susceptible silkworm strain 306 were orally administered with determinate dose of BmNPV. After oral ingestion, midgut, haemolymph and fat body tissues were collected at different time to extract DNA. Fluorescence quantitative PCR amplification of BmNPV DNA polymerase gene (dnapol)was used to detect copy number of BmNPV, and each sample was normalized using silkworm cytoplasm actin A3. The results showed that BmNPV could be detected in the midgut 2 hours post infection.  BmNPV could be detected in the haemolymph and fat body 12 hours post infection. Through a latent period of 12 hours, rapid proliferation of virus was detected in the midgut, haemolymph and fat body. The proliferation of BmNPV entered a stationary phase 84 hours post infection.

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Key words: Bombyx mori, nuclear polyhedrosis virus, DNA polymerase gene, actin gene, proliferation, fluorescence quantitative PCR