We report a disposable and highly effective polymeric microfluidic viral sample concentration device capable of increasing the concentration of virus in a human nasopharyngeal specimen more than one order of magnitude in less than 30 min without the use of a centrifuge. The device is fabricated using 3D maskless xurography method using commercially available polymeric materials, which require no cleanroom operations. The disposable components can be fabricated and assembled in five minutes. The device can concentrate a few milliliters (mL) of influenza virus in solution from tissue culture or clinical nasopharyngeal swab specimens, via reduction of the fluid volume, to tens of microliters (mL). The performance of the device was evaluated by nucleic acid extraction from the concentrated samples, followed by a real-time quantitative polymerase chain reaction (qRT-PCR). The viral RNA concentration in each sample was increased on average over 10-fold for both cultured and patient specimens compared to the starting samples, with recovery efficiencies above 60% for all input concentrations. Highly concentrated samples in small fluid volumes can increase the downstream process speed of on-chip nucleic acid extraction, and result in improvements in the sensitivity of many diagnostic platforms that interrogate small sample volumes.
References
[1]
Castellanos, A.; Ramos, A.; Gonzlez, A.; Green, N.G.; Morgan, H. Electrohydrodynamics and dielectrophoresis in microsystems: Scaling laws. J. Phys. D Appl. Phys. 2003, 36, 2584–2597, doi:10.1088/0022-3727/36/20/023.
[2]
Jacoby, G.A.; Han, P. Detection of extended-spectrum beta-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli. J. Clin. Microbiol. 1996, 34, 908–911.
[3]
Jie, W. Biased AC electro-osmosis for on-chip bioparticle processing. IEEE Trans. Nanotechnol. 2006, 5, 84–89, doi:10.1109/TNANO.2006.869645.
[4]
Ocvirk, G.; Munroe, M.; Tang, T.; Oleschuk, R.; Westra, K.; Harrison, D.J. Electrokinetic control of fluid flow in native poly(dimethylsiloxane) capillary electrophoresis devices. Electrophoresis 2000, 21, 107–115, doi:10.1002/(SICI)1522-2683(20000101)21:1<107::AID-ELPS107>3.0.CO;2-Y.
[5]
Boguslaw, B.; Szumski, M.; Klodzinska, E.; Dahm, H. Separation of bacteria by capillary electrophoresis. J. Separ. Sci. 2003, 26, 1045–1049, doi:10.1002/jssc.200301442.
[6]
Lay, C.; Teo, C.Y.; Zhu, L.; Peh, X.L.; Ji, H.M.; Chew, B.-R.; Murthy, R.; Feng, H.H.; Liu, W.-T. Enhanced microfiltration devices configured with hydrodynamic trapping and a rain drop bypass filtering architecture for microbial cells detection. Lab Chip 2008, 8, 830–833, doi:10.1039/b800015h.
[7]
Zhu, L.; Zhang, Q.; Feng, H.; Ang, S.; Chau, F.S.; Liu, W.-T. Filter-based microfluidic device as a platform for immunofluorescent assay of microbial cells. Lab Chip 2004, 4, 337–341, doi:10.1039/b401834f.
[8]
Reichmuth, D.S.; Wang, S.K.; Barrett, L.M.; Throckmorton, D.J.; Einfeld, W.; Singh, A.K. Rapid microchip-based electrophoretic immunoassays for the detection of swine influenza virus. Lab Chip 2008, 8, 1319–1324, doi:10.1039/b801396a.
[9]
Liu, R.H.; Yang, J.; Lenigk, R.; Bonanno, J.; Grodzinski, P. Self-contained, fully integrated biochip for sample preparation, polymerase chain reaction amplification, and DNA microarray detection. Anal. Chem. 2004, 76, 1824–1831, doi:10.1021/ac0353029.
[10]
Furdui, V.I.; Harrison, D.J. Immunomagnetic T cell capture from blood for PCR analysis using microfluidic systems. Lab Chip 2004, 4, 614–618, doi:10.1039/b409366f.
[11]
Lien, K.-Y.; Lin, J.-L.; Liu, C.-Y.; Lei, H.-Y.; Lee, G.-B. Purification and enrichment of virus samples utilizing magnetic beads on a microfluidic system. Lab Chip 2007, 7, 868–875, doi:10.1039/b700516d.
[12]
Berthier, E.; Warrick, J.; Yu, H.; Beebe, D.J. Managing evaporation for more robust microscale assays Part 1. Volume loss in high throughput assays. Lab Chip 2008, 8, 852–859, doi:10.1039/b717422e.
[13]
Berthier, E.; Warrick, J.; Yu, H.; Beebe, D.J. Managing evaporation for more robust microscale assays Part 2. Characterization of convection and diffusion for cell biology. Lab Chip 2008, 8, 860–864, doi:10.1039/b717423c.
[14]
Sharma, N.R.; Lukyanov, A.; Bardell, R.L.; Seifried, L.; Shen, M. Development of an evaporation-based microfluidic sample concentrator. Proc. SPIE 2008, 6886, doi:10.1117/12.764100.
[15]
Timmer, B.H.; van Delft, K.M.; Olthuis, W.; Bergveld, P.; van den Berg, A. Micro-evaporation electrolyte concentrator. Sens. Actuator. BChem. 2003, 91, 342–346.
Zimmermann, M.; Bentley, S.; Schmid, H.; Hunziker, P.; Delamarche, E. Continuous flow in open microfluidics using controlled evaporation. Lab Chip 2005, 5, 1355–1359, doi:10.1039/b510044e.
[18]
Zhang, J.Y.; Do, J.; Premasiri, W.R.; Ziegler, L.D.; Klapperich, C.M. Rapid point-of-care concentration of bacteria in a disposable microfluidic device using meniscus dragging effect. Lab Chip 2010, 10, 3265–3270, doi:10.1039/c0lc00051e.
[19]
Leng, J.; Lonetti, B.; Tabeling, P.; Joanicot, M.; Ajdari, A. Microevaporators for kinetic exploration of phase diagrams. Phys. Rev. Lett. 2006, 96, 084503:1–084503:4.
[20]
Wang, F.; Chon, C.H.; Li, D. Particle separation by a moving air-liquid interface in a microchannel. J. Colloid Interface Sci. 2010, 352, 580–584, doi:10.1016/j.jcis.2010.08.062.
[21]
Wong, T.-S.; Chen, T.-H.; Shen, X.; Ho, C.-M. Nanochromatography driven by the coffee ring effect. Anal. Chem. 2011, 83, 1871–1873.
[22]
Deegan, R.D.; Bakajin, O.; Dupont, T.F.; Huber, G.; Nagel, S.R.; Witten, T.A. Capillary flow as the cause of ring stains from dried liquid drops. Nature 1997, 389, 827–829.
[23]
Hu, H.; Larson, R.G. Marangoni effect reverses coffee-ring depositions. J. Phys. Chem. B 2006, 110, 7090–7094, doi:10.1021/jp0609232.
[24]
Schaldach, C.M.; Bourcier, W.L.; Shaw, H.F.; Viani, B.E.; Wilson, W.D. The influence of ionic strength on the interaction of viruses with charged surfaces under environmental conditions. J. Colloid Interface Sci. 2006, 294, 1–10, doi:10.1016/j.jcis.2005.06.082.
[25]
Kirby, B.J.; Hasselbrink, E.F. Zeta potential of microfluidic substrates: 2. Data for polymers. Eelectrophoresis 2004, 25, 203–213, doi:10.1002/elps.200305755.
[26]
World Health Organization (WHO). WHO Manual on Animal Influenza Diagnosis and Surveillance. Available online: http://www.wpro.who.int/emerging_diseases/documents/docs/manualonanimalaidiagnosisandsurveillance.pdf (accessed on 18 January 2013).
[27]
Haldar, J.; Weight, A.K.; Klibanov, A.M. Preparation, application and testing of permanent antibacterial and antiviral coatings. Nat. Protocol. 2007, 2, 2412–2417, doi:10.1038/nprot.2007.353.
[28]
CDC. CDC Protocol of Realtime RTPCR for Influenza A(H1N1). Available online: http://www.who.int/entity/csr/resources/publications/swineflu/CDCRealtimeRTPCR_SwineH1Assay-2009_20090430.pdf (accessed on 18 January 2013).