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Rapid Diagnosis of Dengue Viremia by Reverse Transcriptase-Polymerase Chain Reaction using 3′-Noncoding Region Universal Primers
T. Mirawati Sudiro
T. Mirawati Sudiro
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Hiroaki Ishiko
Hiroaki Ishiko
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Sharone Green
Sharone Green
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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David W. Vaughn
David W. Vaughn
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Ananda Nisalak
Ananda Nisalak
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Siripen Kalayanarooj
Siripen Kalayanarooj
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Alan L. Rothman
Alan L. Rothman
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Boonyos Raengsakulrach
Boonyos Raengsakulrach
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Jurand Janus
Jurand Janus
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Ichiro Kurane
Ichiro Kurane
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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Francis A. Ennis
Francis A. Ennis
Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Department of Virology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences, Department of Infectious Disease, Bangkok Children's Hospital, Worcester, Massachusetts, Thailand
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A reverse transcriptase-polymerase chain reaction (RT-PCR) method was developed as a rapid diagnostic test of dengue viremia. To detect dengue viruses in serum or plasma specimens, a pair of universal primers was designed for use in the RT-PCR. Using these primers, the 3′-noncoding region of dengue virus types 1, 2, 3, and 4 could be amplified, but not those of other flaviviruses, such as West Nile virus, Japanese encephalitis virus, and yellow fever virus, or the alphavirus Sindbis virus. The sensitivity of the RT-PCR assay was similar to that of a quantitative fluorescent focus assay of dengue viruses in cell culture. Combining a silica method for RNA isolation and RT-PCR dengue virus could be detected in a 6-hr assay. In a preliminary study using this method, we detected dengue virus in 38 of 39 plasma specimens from which dengue virus had been isolated by mosquito inoculation. We then applied this method for detecting dengue viremia to 117 plasma samples from 62 children with acute febrile illnesses in a dengue-endemic area. We detected dengue viremia in 19 of 20 samples obtained on the day of presentation, which had been confirmed as acute dengue infection by mosquito inoculation and antibody responses. The overall sensitivity of this method was 91.4% (32 of 35; 95% confidence interval [CI] = 82.2–100%). The results from testing plasma samples from febrile nondengue patients showed a specificity of 95.4% (42 of 44; 95% CI = 89.3–100%).
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| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 2047 | 1155 | 316 |
| Full Text Views | 42 | 5 | 0 |
| PDF Downloads | 45 | 7 | 0 |