Amplification and Sequencing of Densonucleosis Virus DNA in C6/36 Cell Line of Aedes albopictus Infected by Strain NGC of Dengue Virus Type 2
［Key words］ dengue virus; Aedes; densonucleosis virus in C6/36 cell of Aedes albopictus; reverse transcriptase polymerase chain reaction; DNA order
To study the reason of insensitiveness of C6/36 cell line in Aedes albopictus to type 2 of dengue virus (DEN2). Methods: The primers were designed according to the sequence of NS1 gene of dengue virus strain NGC. Three groups of C6/36 cell line were set up: virus control group (group A), virus DNA enzyme (DNase) treated group (group B), and cell control group (group C). Nucleic acid of C6/36 cell line was isolated, and RT-PCR, restriction enzyme（Hind Ⅲ）digestion, electrophoresis analysis with 0.8% agarosegels were carried out on it.The retrieval products were sequenced. The homology of the obtained nucleotide sequences with the Densonucleosis virus (DNV）in C6/36 cell line of Aedes albopictus was compared using the PHYLIP software. Results: A band of about 1 300 bp appeared in groups A and B, and it couldn't be cut by Hind Ⅲ. This band could not beamplified from group B. The comparison analysis result showed that the obtained nucleotide sequence had a homology of 95% with part of DNV sequence of C6/36 in Aedes albopictus. Conclusions: Aedes albopictus densonucleosis virus contamination might be the reason for the insensitiveness of C6/36 cell line to type 2 of Dengue virus.
