A portable, integrated analyzer for microfluidic – based molecular analysis
Tóm tắt
A portable, fully automated analyzer that provides actuation and flow control to a disposable, self-contained, microfluidic cassette (“chip”) for point-of-care, molecular testing is described. The analyzer provides mechanical actuation to compress pouches that pump liquids in the cassette, to open and close diaphragm valves for flow control, and to induce vibrations that enhance stirring. The analyzer also provides thermal actuation for the temperature cycling needed for polymerase chain reaction (PCR) amplification of nucleic acids and for various drying processes. To improve the temperature uniformity of the PCR chamber, the system utilizes a double-sided heating/cooling scheme with a custom feedforward, variable, structural proportional-integral-derivative (FVSPID) controller. The analyzer includes a programmable central processing unit that directs the sequence and timing of the various operations and that is interfaced with a computer. The disposable cassette receives a sample, and it carries out cell lysis, nucleic acid isolation, concentration, and purification, thermal cycling, and either real time or lateral flow (LF) based detection. The system’s operation was demonstrated by processing saliva samples spiked with B. cereus cells. The amplicons were detected with a lateral flow assay using upconverting phosphor reporter particles. This system is particularly suited for use in regions lacking centralized laboratory facilities and skilled personnel.
Tài liệu tham khảo
R.H. Birkhahn, E. Haines, W. Wen, L. Reddy, W.M. Briggs, P.A. Datillo, Estimating the clinical impact of bringing a multimarker cardiac panel to the bedside in the ED. Am. J. Emerg. Med. 29, 304 (2011)
R. Boom, C.J. Sol, M.M. Salimans, C.L. Jansen, P.M. Wertheim-van Dillen, J.V.D. Noordaa, Rapid and simple method for purification of nucleic acids. J. Clin. Microbiol. 28, 495 (1990)
Z. Chen, M.G. Mauk, J. Wang, W.R. Abrams, P.L.A.M. Corstjens, R.S. Niedbala, D. Malamud, H.H. Bau, A microfluidic system for saliva-based detection of infectious diseases. Ann. NY Acad. Sci. 1098, 429 (2007)
D. Chen, M.G. Mauk, X. Qiu, C. Liu, J. Kim, S. Ramprasad, S. Ongagna, W.R. Abrams, D. Malamud, P.L.A.M. Corstjens, H.H. Bau, An integrated microfluidic cassette for isolation, amplification, and detection of nucleic acids. Biomed. Microdevices 12, 705 (2010)
C.J. Easley, J.M. Karlinsey, J.M. Bienvenue, L.A. Legendre, M.G. Roper, S.H. Feldman, M.A. Hughes, E.L. Hewlett, T.J. Merkel, J.P. Ferrance, J.P. Landers, A fully integrated microfluidic genetic analysis system with sample-in–answer-out capability. PNAS 103, 19272 (2006)
J. Kim, D. Byun, M.G. Mauk, H.H. Bau, A disposable, self-contained PCR chip. Lab Chip 9, 606 (2009)
E.T. Lagally, J.R. Scherer, R.G. Blazerj, N.M. Toriello, B.A. Diep, M. Ramchandani, G.F. Sensabaugh, L.W. Riley, R.A. Mathies, Integrated portable genetic analysis microsystem for pathogen/infectious disease detection. Anal. Chem. 76, 3162 (2004)
E. Lee-Lewandrowski, K. Lewandrowski, Perspectives on cost and outcomes for point-of-care testing. Clin. Lab. Med. 29, 479 (2009)
R.H. Liu, J. Yang, R. Lenigk, J. Bonanno, P. Grodzinski, Self-contained, fully integrated biochip for sample preparation, polymerase chain reaction, amplification, and DNA microarray detection. Anal. Chem. 76, 1824 (2004)
J.S. Marcus, W.F. Anderson, S.R. Quake, Parallel picoliter RT-PCR assays using microfluidics. Anal. Chem. 78, 956 (2006)
K. Pothier, M. Kirtland, S. Gupta, Has point-of-care come of age? POC 9, 147 (2010)
X. Qiu, J. Yuan, Z. Wang, Chin. J. Chem. Eng. 14, 200 (2006)
X. Qiu, J.A. Thompson, Z. Chen, C. Liu, D. Chen, S. Ramprasad, M.G. Mauk, S. Ongagna, C. Barber, W.R. Abrams, D. Malamud, P.L.A.M. Corstjens, H.H. Bau, Finger-actuated, self-contained immunoassay cassettes. Biomed. Microdevices 11, 1175 (2009)
X. Qiu, M.G. Mauk, D. Chen, C. Liu, H.H. Bau, A large volume, portable, real-time PCR reactor. Lab Chip 10, 3170 (2010)
X. Qiu, C. Liu, M.G. Mauk, R.W. Hart, D. Chen, J. Qiu, T. Kientz, J. Fiene, H.H. Bau, A portable analyzer for pouch-actuated, immunoassay cassettes. Sens. Actuators B. submitted (2011)
F.V.D. Rijke, H. Zijlmans, S. Li, T. Vail, A.K. Raap, R.S. Niedbala, H.J. Tanke, Up-converting phosphor reporters for nucleic acid microarrays. Nat. Biotech. 19, 273 (2001)
M.G. Roper, C.J. Easley, J.P. Landers, Advances in polymerase chain reaction on microfluidic chips. Anal. Chem. 77, 3887 (2005)
J. Wang, Z. Chen, P.L.A.M. Corstjens, M.G. Mauk, H.H. Bau, A disposable microfluidic cassette for DNA amplification and detection. Lab Chip 6, 46 (2006)
C. Zhang, J. Xu, W. Ma, W. Zheng, PCR microfluidic devices for DNA amplification. Biotech. Advan. 24, 243 (2006)