Amplification of complete genome of dengue 2 virus by long RT-PCR

  Objective   To establish the long reverse transcription PCR for amplification of the complete genome of dengue 2 virus.

  Methods   The primers were designed according to the published nucleotide sequence of DEN2NGC strain.Sp6 was added to 5.terminal and restriction endonuclease site for Cla ⅳ was added to 3' terminal.After virus RNA was extracted from the brains of the infected new-born mice, the co mplete genome of DEN2NGC strains was amplified by the long RT-PCR.Three fragments of specific lengths were re-amplified from PCR products for identification.

  Results   The 11kb full-length genome of dengue 2 virus was amplified successfully by the long RT-PCR.

  Conclusion   The complete genome of dengue 2 virus was successful to be amplified using the long RT-PCR, and it will be advantageous for further constructing infectious clone.