Performance of CDC Trioplex qPCR during a dengue outbreak in Brazil

Author(s)

Colombo, Tatiana Elias ^{[1, 2]} ; Versiani, Alice Freitas ^[1] ; Dutra, Karina Rocha ^[1] ; Dias Rubiato, Julia Guimaraes ^[2] ; Galvao, Tayna Manfrin ^[2] ; Negri Reis, Andreia Francesli ^[3] ; Nogueira, Mauricio Lacerda ^[1]

Affiliation

^[1] Fac Med Sao Jose do Rio Preto FAMERP, Sao Jose Do Rio Preto, SP - Brazil

^[2] Univ Paulista UNIP, Sao Jose Do Rio Preto, SP - Brazil

^[3] Prefeitura Sao Jose do Rio Preto, Dept Vigilancia Saude, Sao Jose Do Rio Preto, SP - Brazil

• Document type: Journal article
• Source: Journal of Clinical Virology; v. 121, DEC 2019.
• Web of Science Citations: 0

Abstract

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 (R) 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. (AU)

• FAPESP's process: 18/17647-0 - Development of a peptide-based serologic multiplex diagnostic nanodevices to differentially identify Dengue and Zika infections
• Grantee: Alice Freitas Versiani
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 13/21719-3 - Epidemiological study of dengue (serotypes1-4) in a cohort of São José do Rio Preto, São Paulo, Brazil, during 2014-2018
• Grantee: Maurício Lacerda Nogueira
• Support Opportunities: Research Projects - Thematic Grants