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.

Early and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza viruses at the point-of-care is crucial for reducing disease transmission during the current pandemic and future flu seasons. To prepare for potential cocirculation of these two viruses, we report a valve-enabled, paper-based sample preparation device integrated with isothermal amplification for their simultaneous detection. The device incorporates (1) virus lysis and RNA enrichment, enabled by ball-based valves for sequential delivery of reagents with no pipet requirement, (2) reverse transcription loop-mediated isothermal amplification, carried out in a coffee mug, and (3) colorimetric detection. We have used the device for simultaneously detecting inactivated SARS-CoV-2 and influenza A H1N1 viruses in 50 min, with limits of detection at 2 and 6 genome equivalents, respectively. The device was further demonstrated to detect both viruses in environmental samples.

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Escherichia coli (E. coli) cells are present in fecal materials that can be the main source for disease‐causing agents in water. As a result, E. coli is recommended as a water quality indicator. We have developed an innovative platform to detect E. coli for monitoring water quality on-site by integrating paper-based sample preparation with nucleic acid isothermal amplification. The platform carries out bacterial lysis and DNA enrichment onto a paper pad through ball-based valves for fluid control, with no need of laboratory equipment, followed by loop-mediated isothermal amplification (LAMP) in a battery-operated coffee mug, and colorimetric detection. We have used the platform to detect E. coli in environmental water samples in about 1 h, with a limit of quantitation of 0.2 CFU/mL, and 3 copies per reaction. The platform was confirmed for detecting multiple E. coli strains, and for water samples of different salt concentrations. We validated the functions of the platform by analyzing recreational water samples collected near the Atlantic Ocean that contain different concentrations of salt and bacteria.

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.

Early and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the point-of-care (POC) is crucial for reducing the transmission of coronavirus disease 2019 (COVID-19). To address this need, we have developed a valve-enabled lysis, paper-based RNA enrichment, and RNA amplification device (VLEAD) for detecting SARS-CoV-2. We have combined VLEAD with a smart coffee mug for sample preparation, nucleic acid isothermal amplification, and colorimetric detection using a smartphone camera or a naked eye. VLEAD enables two critical functions required for POC testing: sample preparation and detection. Since the reagents can be pre-packaged in the device, all operational steps can be carried out at POC. We have demonstrated the functions of the device by analyzing samples spiked with heat-inactivated SARS-CoV-2, which were obtained from BEI Resources. This rapid and highly sensitive POC platform for SARS-CoV-2 detection has a potential to help reduce COVID-19 transmission.

We report a point-of-care (POC) device for simultaneous detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A viruses. The device carries out sample preparation using ball-based valves for sequential delivery of reagents. A microfluidic paper-based analytical device (μPAD) in the detection unit enables RNA isolation and enrichment, followed by reverse transcription loop-mediated isothermal amplification (RT-LAMP) and colorimetric detection. The device integrates all the necessary steps for the sample preparation, including virus lysis, RNA enrichment and purification of two virus samples. The device enabled simultaneous detection of SARS-CoV-2 and Influenza A N1H1 viruses in 50 min., with limit of detection of 2 and 6 genome equivalents (GEs), respectively. The device was also capable of detecting environmental sample of the two viruses.

There have been numerous Zika virus (ZIKV) outbreaks in the past few years, representing a public health problem. The recommended tests for the diagnosis of Zika infections are performed in a laboratory setting. However, diagnostics platforms at the point-of-care (POC) are highly desirable for understanding and preventing ZIKV transmission. To address this need, we have developed a testing platform that (1) can be operated in the field for pathogen detection, (2) is rapid, cost-effective, and reliable, and (3) does not require a power supply. To realize the platform, we have developed (1) a series of ball-based valves for the storage and sequential delivery of reagents and (2) microheater-enabled RNA amplification, both of which are integral components of this POC device. The multiple reagents are needed for virus lysis, RNA enrichment and purification. These ball-based are employed for fluid-control and they are actuated manually by sliding the unit and a pole under it, which can lift the balls. Nucleic acid amplification is then performed by a smart coffee mug that provides a constant temperature for reverse transcription loop mediated isothermal amplification (RT-LAMP), followed by colorimetric detection. We have demonstrated the detection of Zika virus in human urine and saliva samples using this testing platform.