title: Sensitivity and specificity of a universal primer set for the rapid diagnosis of dengue virus infections by polymerase chain reaction and nucleic acid hybridization.
abstract: A set of sense and anti-sense oligomeric DNA primers, degenerate in the third "wobble" base position of codons so as to match all known dengue virus sequences, was evaluated as universal primers in a polymerase chain reaction (PCR) assay for the rapid diagnosis of dengue virus infections. Virus-specific complementary DNA (cDNA) was prepared by reverse transcription (RT) of total RNA extracted from serum. Amplified cDNA was identified by nucleic acid hybridization with four serotype-specific, oligomeric DNA probes. Using sera from patients admitted with dengue fever, RT/PCR followed by nucleic acid hybridization using radiolabeled probes was 68% sensitive (50/74; 95% confidence interval [CI] = 57-78%) and 100% specific. Chemiluminescent detection of hybridized products was 62% sensitive (26/42; 95% CI = 46-75%). Using specimens from which a virus isolate had been obtained, RT/PCR followed by nucleic acid hybridization with radiolabeled probes was 80% sensitive (40/50; 95% CI = 69-91%) and 100% specific. The results suggest that RT/PCR using degenerate primers is a sensitive and specific method for the detection of dengue viruses in clinical specimens.
authors.affiliation: Department of Virus Diseases, Walter Reed Army Institute of Research, Washington, DC.
authors.forename: E A
authors.initials: EA
authors.lastname: Henchal
authors.forename: S L
authors.initials: SL
authors.lastname: Polo
authors.forename: V
authors.initials: V
authors.lastname: Vorndam
authors.forename: C
authors.initials: C
authors.lastname: Yaemsiri
authors.forename: B L
authors.initials: BL
authors.lastname: Innis
authors.forename: C H
authors.initials: CH
authors.lastname: Hoke
mesh_terms: D015342:DNA Probes / Q000737:chemistry; D003715:Dengue / Q000175:diagnosis*; D003716:Dengue Virus / Q000235:genetics* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012367:RNA, Viral / Q000097:blood*; D014158:Transcription, Genetic
chemical_list: D015342:DNA Probes; D012367:RNA, Viral