Rapid diagnosis of Dengue viremia by reverse transcriptase-polymerase chain reaction using 3'-noncoding region universal primers

被引:53
作者
Sudiro, TM
Ishiko, H
Green, S
Vaughn, DW
Nisalak, A
Kalayanarooj, S
Rothman, AL
Raengsakulrach, B
Janus, J
Kurane, I
Ennis, FA
机构
[1] ARMED FORCES RES INST MED SCI, DEPT VIROL, USA MED COMPONENT, BANGKOK, THAILAND
[2] BANGKOK CHILDRENS HOSP, DEPT INFECT DIS, BANGKOK, THAILAND
关键词
D O I
10.4269/ajtmh.1997.56.424
中图分类号
R1 [预防医学、卫生学];
学科分类号
1004 ; 120402 ;
摘要
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 at 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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页码:424 / 429
页数:6
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