The performance of RT-PCR compared with a rapid serological assay for acute dengue fever in a diagnostic laboratory

Abstract

The laboratory diagnosis of dengue has largely relied on serological assays, although many different RT-PCR protocols have been reported. Owing to its limited use, the value of RT-PCR in the clinical laboratory has not been fully evaluated. During the outbreak of severe acute respiratory syndrome (SARS) in Singapore in 2003, RT-PCR to detect dengue viral RNA was used as a rapid diagnostic tool to differentiate dengue from SARS among patients who presented to a hospital designated to manage and quarantine SARS cases. A total of 343 results for RT-PCR and 439 results for serology were analysed and compared with the final discharge diagnosis. Our experience indicates that RT-PCR for dengue can be set up rapidly in a clinical laboratory, with very sensitive and specific results for the diagnosis of dengue, particularly in the first 5 days from onset of symptoms.

Cost-Effective Real-Time Reverse Transcriptase PCR (RT-PCR) To Screen for Dengue Virus followed by Rapid Single-Tube Multiplex RT-PCR for Serotyping of the Virus

“…The pan-dengue generic primers used in the present study were a modification of the ones used in previous published study (2). The original primer pair was found to cross-react with West Nile (New York strain), while the specificity of primers in the present study had been demonstrated by tests with closely related flaviviruses.…”

Validation of Probability Equation and Decision Tree in Predicting Subsequent Dengue Hemorrhagic Fever in Adult Dengue Inpatients in Singapore

“…The PCR-positive cases were early presenters during the febrile viremic phase. 22 Although other studies have reported different methods to predict DHF, our study has now validated our predictive tools using different cohorts with different dominant serotypes. Of importance, our predictive tools were intended not to miss a case of DHF, and the high sensitivity and negative predictive value in the validation cohort confirmed the use of the probability equation and decision tool.…”

“…20,21 and confirmed cases had positive dengue polymerase chain reaction (PCR) assay. 22 Dengue hemorrhagic fever was diagnosed when all four criteria of fever, thrombocytopenia ≤ 100 × 10 9 /L, hemorrhagic manifestations, and plasma leakage (hypoproteinemia, ≥ 20% change in hematocrit, or pleural effusion or ascites) were present. 19 Dengue shock syndrome (DSS) is defined as DHF and tachycardia > 100/min or narrow pulse pressure < 20 mmHg, or DHF and systolic blood pressure < 90 mmHg.…”

Dengue Diagnosis: Challenges and Opportunities

“…Combination of RT-PCR and digestion of restriction enzyme of amplified DNAs have been used for the development of a fast, simple virus identification method. Falsepositive results and cross reactivity can be overcome by excluding sequences that are mutual in both the Dengue genome and the DNA or RNA of the human, mosquito [45,46].…”