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Single rapid TaqMan fluorogenic probe based PCR assay that detects all four dengue serotypes
Author(s) -
Warrilow David,
Northill Judith A.,
Pyke Alyssa,
Smith Greg A.
Publication year - 2002
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.2176
Subject(s) - dengue fever , virology , dengue virus , serotype , taqman , flavivirus , japanese encephalitis , polymerase chain reaction , biology , flaviviridae , virus , encephalitis , viral disease , gene , genetics
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. J. Med. Virol. 66:524–528, 2002. © 2002 Wiley‐Liss, Inc.