Performance of CDC Trioplex qPCR during a dengue outbreak in Brazil

Tatiana Elias Colombo, Alice Freitas Versiani, Karina Rocha Dutra, Julia Guimarães Dias Rubiato, Tayna Manfrin Galvão, Andréia Francesli Negri Reis, Maurício Lacerda Nogueira

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


Background: In recent years real‑time reverse transcription polymerase chain reaction (real-time RT-PCR) has become a leading technique for nucleic acid detection and quantification of flaviviruses, including Dengue virus (DENV). Trioplex real-time RT-PCR has the advantages of providing the concurrent detection of Zika virus (ZIKV), DENV, and Chikungunya virus (CHIKV) RNA in human serum. Objective: This study sought to compare the sensitivity and specificity of the Trioplex real-time RT-PCR assay to those provided by CDC DENV TaqMan® RT-qPCR assay and conventional PCR when used for DENV detection in the context of a dengue epidemic. Study design: We analyzed 1656 serum samples from symptomatic patients with acute febrile disease for 5 days less between December 2018 and May 2019. The samples were tested using the various PCR-based assays. Results: Of the 1656 serum samples analyzed, 713 (43%) were laboratory-confirmed as arboviruses: 99.86% (712/713) were confirmed as DENV and 0.14% (1/713) were confirmed as ZIKV. Next, 590 samples were selected, and of these, 331 samples (56.1%) were determined to be positive (Ct < 38) and 259 samples (43.9%) were determined to be negative (Ct > 38) using the Trioplex real-time RT-PCR assay. The multiplex method found that the test exhibits 95% sensitivity and 100% specificity. Conclusion: This evaluation demonstrates the capacity of the Trioplex real-time RT-PCR assay to detect DENV at a high sensitivity and specificity in a geographic area with a current dengue outbreak and a lower co-circulation of other arboviruses – such as ZIKV and CHIKV, and the results prove it´s applicability as clinical screening test that can serve as a confirmatory test.

Original languageEnglish (US)
Article number104208
JournalJournal of Clinical Virology
StatePublished - Dec 2019
Externally publishedYes


  • Dengue virus
  • Detection
  • Sensitivity
  • Specificity

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases


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