Single rapid TaqMan fluorogenic probe based PCR assay that detects all four dengue serotypes.
Public health laboratories require rapid diagnosis of dengue outbreaks for application of measures such as vector control. We have developed a rapid single fluorogenic probe-based polymerase chain reaction assay for the detection of all four dengue serotypes (FUDRT-PCR). The method employs primers and probe that are complementary to the evolutionarily conserved 3' untranslated region of the dengue genome. The assay detected viral RNA of strains of all four dengue serotypes but not of the flaviviruses Japanese encephalitis virus, Murray Valley encephalitis virus, Kunjin, Stratford, West Nile, Alfuy or Yellow fever. When compared to an existing nested-PCR assay for the detection of dengue on clinical samples, FUDRT-PCR detected dengue 1 (100%, n=14), dengue 2 (85%, n=13), dengue 3 (64%, n=14) and dengue 4 (100%, n=3) with the indicated sensitivities. FUDRT-PCR enables diagnosis of acute dengue infection in four hours from sample receipt. In addition, a single-test procedure should result in a reduction in the number of tests performed with considerable cost savings for diagnostic laboratories.
Warrilow
David
D
Queensland Health Scientific Services, Queensland, Australia. David_Warrilow@health.qld.gov.au
Northill
Judith A
JA
Pyke
Alyssa
A
Smith
Greg A
GA
D020413:3' Untranslated Regions / Q000235:genetics; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D005456:Fluorescent Dyes; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000097:blood; D012680:Sensitivity and Specificity; D012703:Serotyping; D019914:Taq Polymerase / Q000378:metabolism*; D013997:Time Factors
D020413:3' Untranslated Regions; D005456:Fluorescent Dyes; D012367:RNA, Viral; D019914:Taq Polymerase