Fusion PCR for amplification of the full-length cDNA of dengue virus type 2 isolated in China / 军事医学科学院院刊

ABSTRACT

Objective:To establish fusion PCR for amplification of the full-length cDNA of dengue virus type 2. Methods:According to the published nucleotide sequence of D2-43,the primers were devised and the 5′ and 3′ half genomic cDNAs of dengue virus type 2 were amplified by long reverse transcription PCR. Using the PCR products as model,the approximate 11 kb full-length cDNA was amplified by fusion PCR. The sequence containing the 5′ noncoding region was determined by PRISMTM ABI 377 automated sequencer.Results:Using fusion PCR,the full-length cDNA of dengue virus type 2 was successfully amplified and its correctness was proved by partial nucleotide sequences analysis. To our best knowledge, this is the first report of the same kind.Conclusion:Fusion PCR is an effective method to amplify the genomic cDNA of dengue virus.