Sensitivity and Specificity of a Universal Primer Set for the Rapid Diagnosis of Dengue Virus Infections by Polymerase Chain Reaction and Nucleic Acid Hybridization

Erik A. Henchal Department of Virus Diseases, Walter Reed Army Institute of Research, Centers for Disease Control, Department of Virology, Armed Forces Research Institute of the Medical Sciences, Washington, DC, Puerto Rico

Search for other papers by Erik A. Henchal in
Current site
Google Scholar
PubMed Close
,
Stephanie L. Polo Department of Virus Diseases, Walter Reed Army Institute of Research, Centers for Disease Control, Department of Virology, Armed Forces Research Institute of the Medical Sciences, Washington, DC, Puerto Rico

Search for other papers by Stephanie L. Polo in
Current site
Google Scholar
PubMed Close
,
Vance Vorndam Department of Virus Diseases, Walter Reed Army Institute of Research, Centers for Disease Control, Department of Virology, Armed Forces Research Institute of the Medical Sciences, Washington, DC, Puerto Rico

Search for other papers by Vance Vorndam in
Current site
Google Scholar
PubMed Close
,
Chandhana Yaemsiri Department of Virus Diseases, Walter Reed Army Institute of Research, Centers for Disease Control, Department of Virology, Armed Forces Research Institute of the Medical Sciences, Washington, DC, Puerto Rico

Search for other papers by Chandhana Yaemsiri in
Current site
Google Scholar
PubMed Close
,
Bruce L. Innis Department of Virus Diseases, Walter Reed Army Institute of Research, Centers for Disease Control, Department of Virology, Armed Forces Research Institute of the Medical Sciences, Washington, DC, Puerto Rico

Search for other papers by Bruce L. Innis in
Current site
Google Scholar
PubMed Close
, and
Charles H. Hoke Department of Virus Diseases, Walter Reed Army Institute of Research, Centers for Disease Control, Department of Virology, Armed Forces Research Institute of the Medical Sciences, Washington, DC, Puerto Rico

Search for other papers by Charles H. Hoke in
Current site
Google Scholar
PubMed Close
DOI:
https://doi.org/10.4269/ajtmh.1991.45.418
Volume/Issue:
Volume 45: Issue 4
Page(s):
418–428
Restricted access
Get Permissions

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.

Author Notes

Copyright:
Copyright © 1991 by The American Society of Tropical Medicine and Hygiene 1991
Published Online:
Oct 1991
Cover The American Journal of Tropical Medicine and Hygiene
The American Journal of Tropical Medicine and Hygiene
Print ISSN:
0002-9637
Past two years Past Year Past 30 Days
Abstract Views 1533 670 225
Full Text Views 49 12 0
PDF Downloads 59 12 0
 
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save