Division of Vector Borne Diseases, Dengue Branch, Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases, San Juan, PR, 00920, USA.
Division of Preparedness and Emerging Infections, Centers for Disease Control and Prevention, National Centers for Emerging and Zoonotic Infectious Diseases, Atlanta, GA, 30333, USA.
Nat Commun. 2018 Apr 11;9(1):1391. doi: 10.1038/s41467-018-03772-1.
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 10 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.
寨卡病毒（ZIKV）的出现和传播给有登革热（DENV）和基孔肯雅热（CHIKV）病毒传播地区的 ZIKV 感染诊断带来了挑战。为了便于检测 ZIKV 感染，并将这些感染与 DENV 和 CHIKV 区分开来，我们开发了三重实时 RT-PCR 检测法（Trioplex 检测法）。在这里，我们描述了对该检测法的多重和单重格式进行优化，以适用于各种化学和仪器，从而促进全球标准化和实施。我们评估了所有 Trioplex 模式在血清和全血中检测这三种病原体以及在尿液中检测 ZIKV 的分析性能。三种病毒在不同 RNA 提取方式下的检测限接近每毫升 10 个基因组拷贝当量（GCE/mL）。同时检测每个患者的一种以上标本类型可额外提高 6.4%的诊断灵敏度。总的来说，Trioplex 检测法的高灵敏度证明了该检测法在确定寨卡病例方面的实用性。
