A fluorescent DNA probes specific to the conserved terminal 3'-noncoding
region (nucleotides 10653-10678) of dengue virus and a pair of flanking
primers are designed to formulate a dengue specific fluorogenic polymerase
chain reaction (PCR). Optimal assay conditions with zero background are
disclosed which permit the detection of low levels of dengue virus from
clinical specimens. Dengue virus isolates from different geographic
regions can be universally detected and identified by the fluorogenic
RT-PCR assay. Moreover, the assay is specific for dengue 2 virus and does
not recognize other related flaviviruses, including dengue serotypes Louis
encephalitis, yellow fever, and Kunjin viruses. The ) 3, and 4, Japanese
encephalitis, St. assay also efficiently detected immunocomplexed dengue
viruses. The fluorogenic RT-PCR assay readily detected viremia in sera
collected from individuals ill with dengue fever.
