Valve-Enabled Sample Preparation and RNA Amplification in a Coffee Mug for Zika Virus Detection

Abstract

The recent outbreaks of Zika virus (ZIKV) infection represent a public health challenge. Rapid, cost-effective, and reliable diagnostic tools for ZIKV detection at the point of care (POC) are highly desirable, especially for resource-limited nations. To address the need, we have developed an integrated device to achieve sample-to-answer ZIKV detection. The device features innovative ball-based valves enabling the storage and sequential delivery of reagents for virus lysis and a paper-based unit for RNA enrichment and purification. The paper unit is placed in a commercially available coffee mug that provides a constant temperature for reverse transcription loop-mediated isothermal amplification (RT-LAMP), followed by colorimetric detection by naked eye or a cellphone camera. Using the device, we demonstrated the reproducible detection of ZIKV in human urine and saliva samples.

Keywords: RNA; detection; isothermal amplification; viruses.