*Article* **Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1**

#### **Md Alamgir Kabir 1,2, Hussein Zilouchian 2, Mazhar Sher 1,2 and Waseem Asghar 1,2,3,\***


Received: 10 December 2019; Accepted: 12 January 2020; Published: 14 January 2020

**Abstract:** The Zika virus (ZIKV) is an emerging flavivirus transmitted to humans by *Aedes* mosquitoes that can potentially cause microcephaly, Guillain–Barré Syndrome, and other birth defects. Effective vaccines for Zika have not ye<sup>t</sup> been developed. There is a necessity to establish an easily deployable, high-throughput, low-cost, and disposable point-of-care (POC) diagnostic platform for ZIKV infections. We report here an automated magnetic actuation platform suitable for a POC microfluidic sandwich enzyme-linked immunosorbent assay (ELISA) using antibody-coated superparamagnetic beads. The smartphone integrated immunoassay is developed for colorimetric detection of ZIKV nonstructural protein 1 (NS1) antigen using disposable chips to accommodate the reactions inside the chip in microliter volumes. An in-house-built magnetic actuator platform automatically moves the magnetic beads through different aqueous phases. The assay requires a total of 9 min to automatically control the post-capture washing, horseradish peroxidase (HRP) conjugated secondary antibody probing, washing again, and, finally, color development. By measuring the saturation intensity of the developed color from the smartphone captured video, the presented assay provides high sensitivity with a detection limit of 62.5 ng/mL in whole plasma. These results advocate a grea<sup>t</sup> promise that the platform would be useful for the POC diagnosis of Zika virus infection in patients and can be used in resource-limited settings.

**Keywords:** Zika NS1; point of care; colorimetric; smartphone; microfluidic
