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 ap ublic health challenge.R apid, cost-effective,a nd reliable diagnostic tools for ZIKV detection at the point of care (POC) are highly desirable,e specially for resource-limited nations.T oaddress 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 apaper-based unit for RNAenrichmentand purification. The paper unit is placed in ac ommercially available coffee mug that provides ac onstant temperature for reverse transcription loop-mediated isothermal amplification (RT-LAMP), followed by colorimetric detection by naked eye or ac ellphone camera. Using the device,w ed emonstrated the reproducible detection of ZIKV in human urine and saliva samples.Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

CUT-LAMP: Contamination-Free Loop-Mediated Isothermal Amplification Based on the CRISPR/Cas9 Cleavage

“…Loop-mediated isothermal amplification (LAMP) is an ultrasensitive gene amplification technique that is carried out at a constant temperature without the requirement of a thermal cycler. − Compared with other isothermal amplification techniques, such as nucleic acid sequence-based amplification (NASBA), strand displacement amplification (SDA), recombinase polymerase amplification (RPA), and helicase-dependent amplification (HDA), LAMP can achieve 10 9 -fold amplification efficiency in less than 1 h using a single polymerase at a constant temperature. ,, In addition, LAMP also has advantages of other amplification methods such as simplicity, time saving, and low cost. − …”

Network Diagnostics for Rapid Testing, Mapping and Bioinformatics Analysis of SARS-CoV-2 and Other Human Respiratory Viruses in Pandemic Settings

“…There are already a number of reports of rapid isothermal amplification for COVID-19 diagnosis [80,81]. One system utilizes a hot coffee mug to provide the temperature for the RT-LAMP reaction for colorimetric detection of zika virus (ZIKV) RNA in human urine and saliva samples [82]. The device uses an innovative ball-based valve which enables the storage and sequential delivery of reagents in a paper-based unit within the mug for RNA enrichment and purification.…”