Performance of the Trioplex real-time RT-PCR assay for detection of Zika, dengue, and chikungunya viruses

Gilberto A. Santiago, Jesús Vázquez, Sean Courtney, Katia Y. Matías, Lauren E. Andersen, Candimar Colón, Angela E. Butler, Rebecca Roulo, John Bowzard, Julie M. Villanueva and Jorge L. Muñoz-Jordan ()
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Gilberto A. Santiago: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Jesús Vázquez: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Sean Courtney: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Katia Y. Matías: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Lauren E. Andersen: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Candimar Colón: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Angela E. Butler: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Rebecca Roulo: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
John Bowzard: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Julie M. Villanueva: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases
Jorge L. Muñoz-Jordan: Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases

Nature Communications, 2018, vol. 9, issue 1, 1-10

Abstract: Abstract The emergence and spread of Zika virus (ZIKV) presented a challenge to the diagnosis of ZIKV infections in areas with transmission of dengue (DENV) and chikungunya (CHIKV) viruses. To facilitate detection of ZIKV infections, and differentiate these infections from DENV and CHIKV, we developed the Trioplex real-time RT-PCR assay (Trioplex assay). Here, we describe the optimization of multiplex and singleplex formats of the assay for a variety of chemistries and instruments to facilitate global standardization and implementation. We evaluated the analytical performance of all Trioplex modalities for detection of these three pathogens in serum and whole blood, and for ZIKV in urine. The limit of detection for the three viruses and in different RNA-extraction modalities is near 103 genome copy equivalents per milliliter (GCE/mL). Simultaneous testing of more than one specimen type from each patient provides a 6.4% additional diagnostic sensitivity. Overall, the high sensitivity of the Trioplex assay demonstrates the utility of this assay ascertaining Zika cases.

Date: 2018
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DOI: 10.1038/s41467-018-03772-1

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