Direct sequencing of large flavivirus PCR products for analysis of genome variation and molecular epidemiological investigations.
The polymerase chain reaction (PCR) was used to amplify viral cDNAs from selected regions of dengue genomic RNA by using appropriate 'consensus' primers. DNA amplicons containing the structural genes from all 4 dengue serotypes were prepared and directly sequenced using dengue-virus-specific primers. This method can characterize reliably flavivirus field isolates at the molecular level without extensive virus propagation and molecular cloning, and will be a valuable tool for molecular epidemiological studies.
Lewis
J G
JG
Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, CO 80522.
Chang
G J
GJ
Lanciotti
R S
RS
Trent
D W
DW
D001483:Base Sequence; D004279:DNA, Viral / Q000737:chemistry*; D003716:Dengue Virus / Q000235:genetics*; D004812:Epidemiologic Methods; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics; D012194:RNA-Directed DNA Polymerase
D004279:DNA, Viral; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase