Circle-to-circle amplification coupled with microfluidic affinity chromatography enrichment for in vitro molecular diagnostics of Zika fever and analysis of anti-flaviviral drug efficacy

• DOI: 10.1016/j.snb.2021.129723
• Publication Date: 2021-03-06T01:55:45Z
• Authors: Ruben R.G. Soares; Aleksandra Pettke; Agustín Robles-Re...; Sahar Zeebaree; Sibel Ciftci; Marianna Tampere; Aman Russom; Marjo-Riitta Puum...; Mats Nilsson; Narayanan Madaboosi
• Keywords: 0303 health sciences; 03 medical and health sciences; 3. Good health

Abstract Sensitive viral diagnostic methods are increasingly in demand to tackle emerging epidemics. The Zika virus (ZIKV) is particularly relevant in tropical resource limited settings (RLS) and is associated with intermittent epidemics such as the recent 2016 ZIKV outbreak in South America, wherein Zika fever was classified by WHO as a public health emergency of international concern. Thus, there is an urgent need for widespread Zika fever diagnostics and efficient drug therapies. ZIKV diagnostics are typically performed using RT-qPCR in centralized laboratories. While extremely sensitive, RT-qPCR requires rapid heating-cooling cycles, combined with continuous fluorescence measurements to allow quantification, implying high costs and limiting availability of molecular diagnostics in RLS. Here, we report isothermal amplification of ZIKV cDNA using padlock probes followed by two rounds of Rolling Circle Amplification (RCA), termed as circle-to-circle amplification (C2CA), combined with a microfluidic affinity chromatography enrichment (μACE) platform. This platform allowed the detection of