Hands-free smartphone-based diagnostics for simultaneous detection of Zika, Chikungunya, and Dengue at point-of-care
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Infectious diseases remain the world’s top contributors to death and disability, and, with recent outbreaks of Zika virus infections there has been an urgency for simple, sensitive and easily translatable point-of-care tests. Here we demonstrate a novel point-of-care platform to diagnose infectious diseases from whole blood samples. A microfluidic platform performs minimal sample processing in a user-friendly diagnostics card followed by real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) on the same card with pre-dried primers specific to viral targets. Our point-of-care platform uses a commercial smartphone to acquire real-time images of the amplification reaction and displays a visual read-out of the assay. We apply this system to detect closely related Zika, Dengue (types 1 and 3) and Chikungunya virus infections from whole blood on the same pre-printed chip with high specificity and clinically relevant sensitivity. Limit of detection of 1.56e5 PFU/mL of Zika virus from whole blood was achieved through our platform. With the ability to quantitate the target nucleic acid, this platform can also perform point-of-care patient surveillance for pathogen load or select biomarkers in whole blood.
KeywordsLoop mediated isothermal amplification Point-of-care diagnostics Zika diagnostics Smartphone diagnostics Hands-free microfluidics
We thank the staff at the Micro and Nanotechnology Laboratory at UIUC for facilitating the chip fabrication. The work was funded by NSF grant 1534126 and the University of Illinois at Urbana-Champaign.
A.G., A.O., G.L.D, B.T.C and R.B conceived the idea and designed the study. A.G. and A.O., designed and performed the experiments. G.L.D and A.B assisted with the experiments and provided intellectual inputs. H.Y. developed the cradle interface for the smartphone and performed the setup characterization experiments. W.C. and F.S developed the fabrication process for the microfluidic chip. A.G developed image processing algorithms to derive fluorescence intensity values from the assay lanes of the microfluidic chip. All wrote and edited the manuscript.
Compliance with ethical standards
The authors declare no competing financial interests.
- K. Curtis, D. Rudolph, I. Nejad, J. Singleton, PLoS One (2012)Google Scholar
- K.A. Curtis, D.L. Rudolph, D. Morrison, D. Guelig, S. Diesburg, D. McAdams, R.A. Burton, P. LaBarre, M. Owen, J. Virol, Methods 237, 132–137 (2016)Google Scholar
- G.L. Damhorst, C. Duarte-Guevara, W. Chen, T. Ghonge, B.T. Cunningham, R. Bashir, Eng. 1, 324–335 (2015)Google Scholar
- O. Faye, O. Faye, A. Dupressoir, M. Weidmann, M. Ndiaye, A. Alpha. 43, 96–101 (2008)Google Scholar
- A. Gourinat, O. O. Connor, E. Calvez, C. Goarant, Emerg. Infect. Dis. 21, 84–86 (2015)Google Scholar
- S. Hu, M. Li, L. Zhong, S. Lu, Z. Liu, J. Pu, J. Wen, BMC Mol. Biol. 1–15 (2015)Google Scholar
- L. Lai, T.H. Lee, L. Tobler, L. Wen, P. Shi, J. Alexander, H. Ewing, M. Busch, Transf. 52, 447–454 (2012)Google Scholar
- R. S. Lanciotti, O. L. Kosoy, J. J. Laven, J. O. Velez, A. J. Lambert, A. J. Johnson, S. M. Stan, M. R. Duffy, 14 (2008)Google Scholar
- R. S. Lanciotti, A. J. Lambert, M. Holodniy, S. Saavedra, C. Castillo, Emerg. Infect. Dis. 22, 2015–2017 (2016)Google Scholar
- D. Lee, Y. Shin, S. Chung, K. S. Hwang, D. S. Yoon, J. H. Lee, Anal. Chem. 88(24), 12272–12278 (2016). doi: 10.1021/doiacs.analchem.6b03460
- J. Lessler, L. H. Chaisson, L. M. Kucirka, Q. Bi, K. Grantz, H. Salje, A. C. Carcelen, C. T. Ott, J. S. Sheffield, N. M. Ferguson, D. A. T. Cummings, C. J. E. Metcalf, I. Rodriguez-Barraquer, Sci. 46(80), 601–604 (2016)Google Scholar
- V. van der Linden, A. Pessoa, W. Dobyns, A.J. Barkovich, H. van der L. Júnior, E.L.R. Filho, E.M. Ribeiro, M. de C. Leal, P.P. de A. Coimbra, M. de F. V. V. Aragão, I. Verçosa, C. Ventura, R.C. Ramos, D.D.C.S. Cruz, M.T. Cordeiro, V.M.R. Mota, M. Dott, C. Hillard, C.A. Moore, MMWR Morb. Mortal. Wkly Rep. 65, 1343–1348 (2016)CrossRefGoogle Scholar
- Y. Lustig, E. Mendelson, N. Paran, S. Melamed, E. Schwartz, 1–4 (2016a)Google Scholar
- Y. Lustig, B. Mannasse, R. Koren, S. Katz-likvornik, 54, 1–16 (2016b)Google Scholar
- K. O. Murray, R. Gorchakov, A. R. Carlson, R. Berry, L. Lai, M. Natrajan, M. N. Garcia, A. Correa, S. M. Patel, K. Aagaard, M. J. Mulligan, Emerg. Infect. Dis. 23, 99–101 (2017)Google Scholar
- D. Musso, C. Roche, T.-X. Nhan, E. Robin, A. Teissier, V.-M. Cao-Lormeau, Detection of Zika virus in saliva, 68, (2015)Google Scholar
- T. Notomi, H. Okayama, H. Masubuchi, T. Yonekawa, K. Watanabe, N. Amino, T. Hase, 28, (2000)Google Scholar
- M. Parida, G. Posadas, S. Inoue, F. Hasebe, K. Morita, 42, 257–263 (2004)Google Scholar
- M. M. Parida, S. R. Santhosh, P. K. Dash, N. K. Tripathi, V. Lakshmi, N. Mamidi, A. Shrivastva, N. Gupta, P. Saxena, J. P. Babu, P. V. L. Rao, J. Clin. Microbiol. 45, 351–357 (2007)Google Scholar
- M. Safavieh, M. K. Kanakasabapathy, F. Tarlan, M. uddin Ahmed, M. Zourob, W. Asghar, H. Shafiee, ACS Biomater. Sci. Eng. (2016). doi: 10.1021/acsbiomaterials.5b00449
- S. Swaminathan, R. Schlaberg, J. Lewis, K. E. Hanson, M. R. Couturier, 1907–1909 (2016)Google Scholar
- J. J. Waggoner, A. Pinsky, J. Clin. Microbiol. 54, 860–867 (2016)Google Scholar
- J. J. Waggoner, L. Gresh, A. Mohamed-hadley, G. Ballesteros, M. Jose, V. Davila, Y. Tellez, M. K. Sahoo, A. Balmaseda, E. Harris, B. A. Pinsky, 22 (2016a)Google Scholar
- X. Wang, F. Yin, Y. Bi, G. Cheng, J. Li, L. Hou, Y. Li, B. Yang, W. Liu, L. Yang, J. Virol, Methods 238, 86–93 (2016)Google Scholar
- N. N. Watkins, U. Hassan, G. Damhorst, H. Ni, A. Vaid, W. Rodriguez, R. Bashir, Sci. Transl. Med. (2013). doi: 10.1126/scitranslmed.3006870