Abstract
Conventional real-time PCR using fluorescence detection requires expensive optical detection systems with fluorescence labeling. To simplify this PCR system, we proposed an electrochemical impedance spectroscopy (EIS) using an interdigitated electrode integrated inside the PCR chip. The electrode makes a direct contact with the PCR sample and does not require any labeling or immobilization pretreatment. The input AC voltage for EIS showed the lowest noise at 100 mV. Electrical impedances in a frequency domain were measured during 30 cycles in the PCR of Escherichia coli genomic DNA region (of length 180 bp, 10 ng/μl). From the analysis of EIS data, the magnitude of imaginary value steadily increased with an increase in the PCR cycles and showed the greatest change rate at 186 Hz. For comparing the quantitative performance with previous researches, the figure of merit (FM) was defined as the ratio of normalized sensitivity (NS) to the normalized root mean square error (NRMSE). The performance of the proposed EIS method is similar to that reported in other studies, and the damage of the sample monitored through electrophoresis by EIS measurement was confirmed to be negligible.
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References
Ahmed MU, Nahar S, Safavieh M, Zourob M (2013) Real-time electrochemical detection of pathogen DNA using electrostatic interaction of a redox probe. Analyst 138:907–915. https://doi.org/10.1039/c2an36153a
Defever T, Druet M, Evrard D, Marchal D, Limoges B (2011) Real-time electrochemical PCR with a DNA intercalating redox probe. Anal Chem 83:1815–1821. https://doi.org/10.1021/ac1033374
Fang TH, Ramalingam N, Dui DX, Ngin TS, Xianting Z, Kuan ATL, Huat EYP, Qing GH (2009) Real-time PCR microfluidic devices with concurrent electrochemical detection. Biosens Bioelectron 24:2131–2136. https://doi.org/10.1016/j.bios.2008.11.009
Fang X, Zhang H, Zhang F, Jing F, Mao H, Jin Q, Zhao J (2012) Real-time monitoring of strand-displacement DNA amplification by a contactless electrochemical microsystem using interdigitated electrodes. Lab Chip 12:3190–3196. https://doi.org/10.1039/c2lc40384f
Foo KL, Kashif M, Tan SJ, Hashim U (2017) An electrochemical DNA biosensor based gold-thiolate conjugation utilizing ruthenium complex [Ru(dppz)2(qtpy)]Cl2. Microsyst Technol 23:1237–1245. https://doi.org/10.1007/s00542-016-2874-7
Gau JJ, Lan EH, Dunn B, Ho CM, Woo JCS (2001) A MEMS based amperometric detector for E. Coli bacteria using self-assembled monolayers. Biosens Bioelectron 16:745–755. https://doi.org/10.1016/S0956-5663(01)00216-0
Jeong S, Lim J, Kim MY, Yeom JH, Cho H, Lee H, Shin YB, Lee JH (2018) Portable low-power thermal cycler with dual thin-film Pt heaters for a polymeric PCR chip. Biomed Microdev 20:14. https://doi.org/10.1007/s10544-018-0257-9
Dr. Kary Banks Mullis biography (2018) http://www.karymullis.com Accessed 4 Oct 2018
Kivlehan F, Mavre F, Talini L, Limoges B, Marchal D (2011) Real-time electrochemical monitoring of isothermal helicase-dependent amplification of nucleic acids. Analyst 136:3635–3642. https://doi.org/10.1039/c1an15289k
Lagally ET, Simpson PC, Mathies RA (2000) Monolithic integrated microfluidic DNA amplification and capillary electrophoresis analysis system. Sens Actuators, B 63:138–146. https://doi.org/10.1016/S0925-4005(00)00350-6
Lee TMH, Carles MC, Hsing IM (2003) Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection. Lap Chip 3:100–105. https://doi.org/10.1039/b300799e
Lee DS, Park SH, Yang H, Chung KH, Yoon TH, Kim SJ, Kim K, Kim YT (2004) Bulk-micromachined submicroliter-volume PCR chip with very rapid thermal response and low power consumption. Lab Chip 4:401–407. https://doi.org/10.1039/B313547K
Luo X, Hsing IM (2009) Electrochemical techniques on sequence-specific PCR amplicon detection for point-of-care applications. Analyst 134:1957–1964. https://doi.org/10.1039/b912653h
Luo X, Xuan F, Hsing IM (2011) Real time electrochemical monitoring of PCR amplicons using electroactive hydrolysis probe. Electrochem Commun 13:742–745. https://doi.org/10.1016/j.elecom.2011.04.027
Ma H, Wallbank RWR, Chaji R, Li J, Suzuki Y, Jiggins C, Nathan A (2013) An impedance-based integrated biosensor for suspended DNA characterization. Sci Rep 3:2730. https://doi.org/10.1038/srep02730
Mahato K, Maurya PK, Chandra P (2018) Fundamentals and commercial aspects of nanobiosensors in point of care clinical diagnostics. 3 Biotech 8:149. https://doi.org/10.1007/s13205-018-1148-8
Nagatani N, Yamanaka K, Saito M, Koketsu R, Sasaki T, Ikuta K, Miyahara T, Tamiya E (2011) Semi-real time electrochemical monitoring for influenza virus RNA by reverse transcription loop-mediated isothermal amplification using a USB powered portable potentiostat. Analyst 136:5143–5150. https://doi.org/10.1039/c1an15638a
Peng HP, Hu Y, Liu P, Deng YN, Wang P, Chen W, Liu AL, Chen YZ, Lin XH (2015) Label-free electrochemical DNA biosensor for rapid detection of mutidrug resistance gene based on Au nanoparticles/toluidine blue–graphene oxide nanocomposites. Sens Actuators, B 207:269–276. https://doi.org/10.1016/j.snb.2014.10.059
Rajapaksha RDAA, Hashim U, Uda MNA, Fernando CAN (2017) Target ssDNA detection of E.coli O157:H7 through electrical based DNA biosensor. Microsyst Technol 23:5771–5780. https://doi.org/10.1007/s00542-017-3498-2
Tabrizi MA, Shamsipur M (2015) A label-free electrochemical DNA biosensor based on covalent immobilization of salmonella DNA sequences on the nanoporous glassy carbon electrode. Biosens Bioelectron 69:100–105. https://doi.org/10.1016/j.bios.2015.02.024
Won BY, Shin S, Baek S, Jung YL, Li T, Shin SC, Cho DY, Lee SB, Park HG (2011) Investigation of the signaling mechanism and verification of the performance of an electrochemical real-time PCR system based on the interaction of methylene blue with DNA. Analyst 136:1573–1579. https://doi.org/10.1039/c0an00695e
Xiang Q, Xu B, Li D (2007) Miniature real time PCR on chip with multi-channel fiber optical fluorescence detection module. Biomed Microdev 9:443–449. https://doi.org/10.1007/s10544-007-9048-4
Yeung SSW, Lee TMH, Hsing IM (2006) Electrochemical real-time polymerase chain reaction. J Am Chem Soc 128:13374–13375. https://doi.org/10.1021/ja065733j
Yun J, Kim J, Lee S, Cho HH, Lee JH (2018) Novel method for the detection of the facial nerve using electricalimpedance spectroscopy during otologic surgery. Sens Actuators, B 261:467–473. https://doi.org/10.1016/j.snb.2018.01.157
Acknowledgements
This work was partially supported by the GIST Research Institute (GRI) Korea in 2018, and the WeMEMS [Grant number WP-EN-18-1].
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Jeong, S., Lim, J., Kim, J. et al. Label-free electrochemical impedance spectroscopy using a micro interdigitated electrode inside a PCR chip for real-time monitoring. Microsyst Technol 25, 3503–3510 (2019). https://doi.org/10.1007/s00542-018-4250-2
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DOI: https://doi.org/10.1007/s00542-018-4250-2