An Aluminum-Based Microfluidic Chip for Polymerase Chain Reaction Diagnosis

Real-time polymerase chain reaction (real-time PCR) tests were successfully conducted in an aluminum-based microfluidic chip developed in this work. The reaction chamber was coated with silicone-modified epoxy resin to isolate the reaction system from metal surfaces, preventing the metal ions from interfering with the reaction process. The patterned aluminum substrate was bonded with a hydroxylated glass mask using silicone sealant at room temperature. The effect of thermal expansion was counteracted by the elasticity of cured silicone. With the heating process closely monitored, real-time PCR testing in reaction chambers proceeded smoothly, and the results show similar quantification cycle values to those of traditional test sets. Scanning electron microscope (SEM) and atomic force microscopy (AFM) images showed that the surface of the reaction chamber was smoothly coated, illustrating the promising coating and isolating properties. Energy-dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), and inductively coupled plasma-optical emission spectrometer (ICP-OES) showed that no metal ions escaped from the metal to the chip surface. Fourier-transform infrared spectroscopy (FTIR) was used to check the surface chemical state before and after tests, and the unchanged infrared absorption peaks indicated the unreacted, antifouling surface. The limit of detection (LOD) of at least two copies can be obtained in this chip.

[1]  Yibo Gao,et al.  Ultrafast PCR Detection of COVID-19 by Using a Microfluidic Chip-Based System , 2022, Bioengineering.

[2]  J. Radolf,et al.  A suite of PCR-LwCas13a assays for detection and genotyping of Treponema pallidum in clinical samples , 2022, Nature Communications.

[3]  J. Huggett,et al.  Monkeypox: another test for PCR , 2022, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[4]  A. Bhagat,et al.  Modular micro-PCR system for the onsite rapid diagnosis of COVID-19 , 2022, Microsystems & nanoengineering.

[5]  Bo Li,et al.  A new dynamic deep learning noise elimination method for chip-based real-time PCR , 2022, Analytical and Bioanalytical Chemistry.

[6]  W. Wen,et al.  A Rapid Digital PCR System with a Pressurized Thermal Cycler , 2021, Micromachines.

[7]  M. Saeb,et al.  Integration of antifouling properties into epoxy coatings: a review , 2021, Journal of Coatings Technology and Research.

[8]  F. Song,et al.  A proof-of-principle study on implementing polymerase chain displacement reaction (PCDR) to improve forensic low-template DNA analysis. , 2021, Forensic science international. Genetics.

[9]  W. Wen,et al.  Lyophilized Ready-to-Use Mix for the Real-Time Polymerase Chain Reaction Diagnosis , 2021 .

[10]  W. Wen,et al.  Point-of-care testing detection methods for COVID-19. , 2021, Lab on a chip.

[11]  Pouria Fattahi,et al.  A microfluidic platform for cultivating ovarian cancer spheroids and testing their responses to chemotherapies , 2020, Microsystems & nanoengineering.

[12]  R. Zengerle,et al.  Fusing MEMS technology with lab-on-chip: nanoliter-scale silicon microcavity arrays for digital DNA quantification and multiplex testing , 2020, Microsystems & Nanoengineering.

[13]  Amene Saghazadeh,et al.  Microfluidic devices for detection of RNA viruses , 2020, Reviews in medical virology.

[14]  Sheng Zhang,et al.  Profile of RT-PCR for SARS-CoV-2: A Preliminary Study From 56 COVID-19 Patients , 2020, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[15]  D. Acierno,et al.  Thermal Conductivity of Polypropylene-Based Materials , 2019, Polypropylene - Polymerization and Characterization of Mechanical and Thermal Properties.

[16]  Lijuan Zhang,et al.  A fully portable microchip real‐time polymerase chain reaction for rapid detection of pathogen , 2019, Electrophoresis.

[17]  Kuangwen Hsieh,et al.  Programmable microfluidic genotyping of plant DNA samples for marker-assisted selection , 2018, Microsystems & Nanoengineering.

[18]  M. Machovský,et al.  Enhanced Charpy impact strength of epoxy resin modified with vinyl‐terminated polydimethylsiloxane , 2018 .

[19]  Jing Cheng,et al.  Enclosed casting of epoxy resin for rapid fabrication of rigid microfluidic chips , 2017 .

[20]  Andreas Manz,et al.  Polymerase chain reaction in microfluidic devices. , 2016, Lab on a chip.

[21]  M. Schibler,et al.  Ebola virus disease diagnosis by real-time RT-PCR: A comparative study of 11 different procedures. , 2016, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[22]  Chenguang Zhou,et al.  The preparation and properties study of polydimethylsiloxane-based coatings modified by epoxy resin , 2016, Journal of Polymer Research.

[23]  W Hampton Henley,et al.  A microfluidic chip integrating DNA extraction and real-time PCR for the detection of bacteria in saliva. , 2013, Lab on a chip.

[24]  Wei Jin,et al.  Self-priming compartmentalization digital LAMP for point-of-care. , 2012, Lab on a chip.

[25]  Weijia Wen,et al.  Fast detection of genetic information by an optimized PCR in an interchangeable chip , 2012, Biomedical microdevices.

[26]  A. Jannesari,et al.  Effect of Molecular Weight and Content of PDMS on Morphology and Properties of Silicone-Modified Epoxy Resin , 2012 .

[27]  Xiuqing Gong,et al.  Inhibitory effect of common microfluidic materials on PCR outcome , 2012 .

[28]  Mieke Uyttendaele,et al.  The challenge of merging food safety diagnostic needs with quantitative PCR platforms , 2011 .

[29]  D. Meldrum,et al.  Real-time PCR of single bacterial cells on an array of adhering droplets. , 2011, Lab on a chip.

[30]  Nicole A. Leal,et al.  Artificial genetic systems: self-avoiding DNA in PCR and multiplexed PCR. , 2010, Angewandte Chemie.

[31]  Jerry Y. H. Fuh,et al.  Micro-spike EEG electrode and the vacuum-casting technology for mass production , 2009 .

[32]  V. Beneš,et al.  The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. , 2009, Clinical chemistry.

[33]  Doyoung Byun,et al.  A disposable, self-contained PCR chip. , 2009, Lab on a chip.

[34]  F. Monzon.  Replacing PCR with COLD-PCR enriches variant DNA sequences and redefines the sensitivity of genetic testing , 2009 .

[35]  Yunzhi Yang,et al.  The effect of titanium surface roughening on protein absorption, cell attachment, and cell spreading. , 2008, The International journal of oral & maxillofacial implants.

[36]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[37]  J. Halgunset,et al.  Low copy number DNA template can render polymerase chain reaction error prone in a sequence-dependent manner. , 2005, The Journal of molecular diagnostics : JMD.

[38]  Gwo-Bin Lee,et al.  Micromachined polymerase chain reaction system for multiple DNA amplification of upper respiratory tract infectious diseases. , 2005, Biosensors & bioelectronics.

[39]  H. Terryn,et al.  Improving the adhesion between epoxy coatings and aluminium substrates , 2004 .

[40]  S. Reid,et al.  Detection of Trypanosoma evansi in camels using PCR and CATT/T. evansi tests in Kenya. , 2004, Veterinary parasitology.

[41]  P. Gill,et al.  Encoded evidence: DNA in forensic analysis , 2004, Nature Reviews Genetics.

[42]  Carl T Wittwer,et al.  Real-time PCR technology for cancer diagnostics. , 2002, Clinical chemistry.

[43]  A. Nitsche,et al.  Real-time PCR in virology. , 2002, Nucleic acids research.

[44]  Thomas D. Schmittgen,et al.  Real-Time Quantitative PCR , 2002 .

[45]  P E Klapper,et al.  Multiplex PCR: Optimization and Application in Diagnostic Virology , 2000, Clinical Microbiology Reviews.

[46]  V. Mišković‐Stanković,et al.  Corrosion protection of aluminium by a cataphoretic epoxy coating , 1999 .

[47]  A Manz,et al.  Chemical amplification: continuous-flow PCR on a chip. , 1998, Science.

[48]  B. Schaefer,et al.  Revolutions in rapid amplification of cDNA ends: new strategies for polymerase chain reaction cloning of full-length cDNA ends. , 1995, Analytical biochemistry.

[49]  L J Kricka,et al.  PCR in a silicon microstructure. , 1994, Clinical chemistry.

[50]  Russell Higuchi,et al.  Kinetic PCR Analysis: Real-time Monitoring of DNA Amplification Reactions , 1993, Bio/Technology.

[51]  M. McClelland,et al.  Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. , 1991, Nucleic acids research.

[52]  K. Mullis,et al.  Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. , 1988, Science.

[53]  Henry A. Erlich,et al.  Analysis of enzymatically amplified β-globin and HLA-DQα DNA with allele-specific oligonucleotide probes , 1986, Nature.

[54]  K. Mullis,et al.  Specific enzymatic amplification of DNA in vitro: the polymerase chain reaction. , 1986, Cold Spring Harbor symposia on quantitative biology.

[55]  K. Mullis,et al.  Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. , 1985, Science.