{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T17:15:25Z","timestamp":1785258925571,"version":"3.55.0"},"reference-count":30,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,1,14]],"date-time":"2020-01-14T00:00:00Z","timestamp":1578960000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R15AI127214"],"award-info":[{"award-number":["R15AI127214"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R56AI138659"],"award-info":[{"award-number":["R56AI138659"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006827","name":"Florida Department of Health","doi-asserted-by":"publisher","award":["7ZK10"],"award-info":[{"award-number":["7ZK10"]}],"id":[{"id":"10.13039\/100006827","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>The Zika virus (ZIKV) is an emerging flavivirus transmitted to humans by Aedes mosquitoes that can potentially cause microcephaly, Guillain\u2013Barr\u00e9 Syndrome, and other birth defects. Effective vaccines for Zika have not yet 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 great 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.<\/jats:p>","DOI":"10.3390\/diagnostics10010042","type":"journal-article","created":{"date-parts":[[2020,1,15]],"date-time":"2020-01-15T03:20:22Z","timestamp":1579058422000},"page":"42","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1659-3125","authenticated-orcid":false,"given":"Md Alamgir","family":"Kabir","sequence":"first","affiliation":[{"name":"Department of Computer &amp; Electrical Engineering and Computer Science, Florida Atlantic University, Boca Raton, FL 33431, USA"},{"name":"Asghar-Lab, Micro and Nanotechnology in Medicine, College of Engineering and Computer Science, Boca Raton, FL 33431, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hussein","family":"Zilouchian","sequence":"additional","affiliation":[{"name":"Asghar-Lab, Micro and Nanotechnology in Medicine, College of Engineering and Computer Science, Boca Raton, FL 33431, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mazhar","family":"Sher","sequence":"additional","affiliation":[{"name":"Department of Computer &amp; Electrical Engineering and Computer Science, Florida Atlantic University, Boca Raton, FL 33431, USA"},{"name":"Asghar-Lab, Micro and Nanotechnology in Medicine, College of Engineering and Computer Science, Boca Raton, FL 33431, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Waseem","family":"Asghar","sequence":"additional","affiliation":[{"name":"Department of Computer &amp; Electrical Engineering and Computer Science, Florida Atlantic University, Boca Raton, FL 33431, USA"},{"name":"Asghar-Lab, Micro and Nanotechnology in Medicine, College of Engineering and Computer Science, Boca Raton, FL 33431, USA"},{"name":"Department of Biological Sciences (Courtesy Appointment), Florida Atlantic University, Boca Raton, FL 33431, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,1,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3","DOI":"10.4103\/0974-777X.176140","article-title":"The Emergence of Zika Virus as a Global Health Security Threat: A Review and a Consensus Statement of the INDUSEM Joint Working Group (JWG)","volume":"8","author":"Sikka","year":"2016","journal-title":"J. Glob. Infect. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1115\/1.4041086","article-title":"Advances in Diagnostic Methods for Zika Virus Infection","volume":"12","author":"Herrada","year":"2018","journal-title":"J. Med. Devices"},{"key":"ref_3","unstructured":"PAHO, and WHO (2019, October 03). Cases of Zika Virus Disease. Available online: http:\/\/www.paho.org\/data\/index.php\/en\/?option=com_content&view=article&id=524&Itemid=."},{"key":"ref_4","unstructured":"WHO (2019, October 08). Zika Epidemiology Update. Available online: https:\/\/www.who.int\/emergencies\/diseases\/zika\/zika-epidemiology-update-july-2019.pdf?ua=1."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"359","DOI":"10.3201\/eid2102.141363","article-title":"Potential sexual transmission of Zika virus","volume":"21","author":"Musso","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Zhang, L., Du, X., Chen, C., Chen, Z., Zhang, L., Han, Q., Xia, X., Song, Y., and Zhang, J. (2018). Development and characterization of double-antibody sandwich ELISA for detection of zika virus infection. Viruses, 10.","DOI":"10.3390\/v10110634"},{"key":"ref_7","unstructured":"World Health Organization (2016). Laboratory Testing for Zika Virus Infection: Interim Guidance, World Health Organization."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"60","DOI":"10.5858\/arpa.2016-0406-SA","article-title":"Laboratory diagnosis of Zika virus infection","volume":"141","author":"Landry","year":"2016","journal-title":"Arch. Pathol. Lab. Med"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1691","DOI":"10.3201\/eid2209.160725","article-title":"Specificity of dengue NS1 antigen in differential diagnosis of dengue and Zika virus infection","volume":"22","author":"Matheus","year":"2016","journal-title":"Emerg. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"12743","DOI":"10.1021\/acs.analchem.7b02862","article-title":"Aptamer-based ELISA assay for highly specific and sensitive detection of Zika NS1 protein","volume":"89","author":"Lee","year":"2017","journal-title":"Anal. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/j.aca.2018.12.043","article-title":"Smartphone-based fluorescent lateral flow immunoassay platform for highly sensitive point-of-care detection of Zika virus nonstructural protein 1","volume":"1055","author":"Rong","year":"2019","journal-title":"Anal. Chim. Acta"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1021\/acsinfecdis.7b00110","article-title":"Surface-enhanced raman spectroscopy-based sandwich immunoassays for multiplexed detection of zika and dengue viral biomarkers","volume":"3","author":"Bosch","year":"2017","journal-title":"ACS Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2793","DOI":"10.1128\/JCM.02142-09","article-title":"Evaluation of a commercial SD dengue virus NS1 antigen capture enzyme-linked immunosorbent assay kit for early diagnosis of dengue virus infection","volume":"48","author":"Wang","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1016\/j.bios.2016.08.097","article-title":"The capacitive sensing of NS1 Flavivirus biomarker","volume":"87","author":"Cecchetto","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"16381","DOI":"10.1039\/C5NR04987C","article-title":"A gold immunochromatographic assay for the rapid and simultaneous detection of fifteen \u03b2-lactams","volume":"7","author":"Chen","year":"2015","journal-title":"Nanoscale"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4995","DOI":"10.1021\/ac500540z","article-title":"Multiplex lateral flow immunoassay for mycotoxin determination","volume":"86","author":"Song","year":"2014","journal-title":"Anal. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5537","DOI":"10.1021\/acs.langmuir.5b00592","article-title":"Hierarchical Flowerlike Gold Nanoparticles Labeled Immunochromatography Test Strip for Highly Sensitive Detection of Escherichia coli O157:H7","volume":"31","author":"Zhang","year":"2015","journal-title":"Langmuir"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1600920","DOI":"10.1002\/adhm.201600920","article-title":"Lateral Flow Assay Based on Paper\u2013Hydrogel Hybrid Material for Sensitive Point-of-Care Detection of Dengue Virus","volume":"6","author":"Choi","year":"2017","journal-title":"Adv. Healthc. Mater."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1757","DOI":"10.1007\/s00705-018-3776-z","article-title":"Laser-cut paper-based device for the detection of dengue non-structural NS1 protein and specific IgM in human samples","volume":"163","author":"Theillet","year":"2018","journal-title":"Arch. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"8159","DOI":"10.1039\/C8RA07607C","article-title":"Development of a flow-free magnetic actuation platform for an automated microfluidic ELISA","volume":"9","author":"Coarsey","year":"2019","journal-title":"RSC Adv."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3796","DOI":"10.1038\/srep03796","article-title":"Micro-a-fluidics ELISA for Rapid CD4 Cell Count at the Point-of-Care","volume":"4","author":"Wang","year":"2014","journal-title":"Sci. Rep. Artic."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"50","DOI":"10.2807\/1560-7917.ES.2016.21.50.30426","article-title":"Serodiagnosis of Zika virus (ZIKV) infections by a novel NS1-based ELISA devoid of cross-reactivity with dengue virus antibodies: A multicohort study of assay performance, 2015 to 2016","volume":"21","author":"Steinhagen","year":"2016","journal-title":"Eurosurveillance"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"7474","DOI":"10.1021\/acsomega.9b00499","article-title":"Microfluidic Chip for Detection of Fungal Infections","volume":"4","author":"Asghar","year":"2019","journal-title":"ACS Omega"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"111490","DOI":"10.1016\/j.bios.2019.111490","article-title":"Development of a multiplex fully automated assay for rapid quantification of CD4+ T cells from whole blood","volume":"142","author":"Sher","year":"2019","journal-title":"Biosens. Bioelectron."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3411","DOI":"10.1039\/c1lc20479c","article-title":"Integration of cell phone imaging with microchip ELISA to detect ovarian cancer HE4 biomarker in urine at the point-of-care","volume":"11","author":"Wang","year":"2011","journal-title":"Lab-on-a-Chip"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.snb.2018.11.036","article-title":"Point-of-care colorimetric analysis through smartphone video","volume":"282","author":"Coleman","year":"2019","journal-title":"Sens. Actuators B Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1935","DOI":"10.1039\/C8AN02521E","article-title":"Cell phone based colorimetric analysis for point-of-care settings","volume":"144","author":"Coleman","year":"2019","journal-title":"Analyst"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.bios.2017.08.051","article-title":"Novel graphene-based biosensor for early detection of Zika virus infection","volume":"100","author":"Afsahi","year":"2018","journal-title":"Biosens. Bioelectron."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"eaan1589","DOI":"10.1126\/scitranslmed.aan1589","article-title":"Rapid antigen tests for dengue virus serotypes and Zika virus in patient serum","volume":"9","author":"Bosch","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1038\/nrmicro1555","article-title":"Why do we need quality-assured diagnostic tests for sexually transmitted infections?","volume":"4","author":"Peeling","year":"2006","journal-title":"Nat. Rev. Microbiol."}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/10\/1\/42\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:26:21Z","timestamp":1760365581000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/10\/1\/42"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,1,14]]},"references-count":30,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,1]]}},"alternative-id":["diagnostics10010042"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics10010042","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,1,14]]}}}