A new quantitative RT-PCR method for sensitive detection of dengue virus in serum samples.
In order to detect and identify dengue serotypes in serum samples, we developed a single-step quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assay (referred to as Q-PCR). Sets of primers were selected from the capsid region of the viral genome. Dengue serotypes 1/3 and 2/4 were detected in two separate duplex amplification reactions using specific primers and fluorogenic TaqMan probes. Results obtained with this Q-PCR and the classical nested RT-PCR (N-PCR) assays were compared using a panel of 97 representative human sera collected from patients in Bangkok, Thailand. It is shown that the Q-PCR is a rapid, sensitive and reproducible tool for the detection and quantitation of the four dengue serotypes in clinical samples, and therefore of great interest for diagnostic use or for large cohort studies.
Sadon
Nadine
N
GlaxoSmithKline Biologicals, Research and Development, Viral Vaccines, Rixensart, Belgium.
Delers
Anne
A
Jarman
Richard G
RG
Klungthong
Chonticha
C
Nisalak
Ananda
A
Gibbons
Robert V
RV
Vassilev
Ventzislav
V
D036022:Capsid Proteins / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D003715:Dengue / Q000175:diagnosis*; D003716:Dengue Virus / Q000145:classification* / Q000302:isolation & purification*; D005456:Fluorescent Dyes; D006801:Humans; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D044967:Serum / Q000821:virology*; D013785:Thailand; D019562:Viral Load / Q000379:methods
D036022:Capsid Proteins; D017931:DNA Primers; D005456:Fluorescent Dyes