Multianalyte Antibiotic Detection on an Electrochemical Microfluidic Platform.

The excessive use of antibiotics in human and veterinary medicine causes the emergence of multidrug resistant bacteria. In this context, the surveillance of many different antibiotics provokes a worldwide challenge. Hence, fast and versatile multianalyte single-use biosensors are of increasing interest for many fields such as medical analysis or environmental and food control. Here we present a microfluidic platform enabling the electrochemical readout of up to eight enzyme-linked assays (ELAs), simultaneously. To demonstrate the applicability of this platform for the surveillance and monitoring of antibiotics, we used highly sensitive biomolecular sensor systems for the simultaneous detection of two commonly employed antibiotic classes tetracycline and streptogramin. Thus, microfluidic channel networks are designed, comprising distinct numbers of immobilization sections with a very low volume of 680 nL each. These passively metered sections can be actuated separately for an individual assay procedure. The limits of detection (LOD) are determined, with high precision, to 6.33 and 9.22 ng mL-1 for tetracycline and pristinamycin, respectively. The employed channel material, dry film photoresist (DFR), allows an easy storage of preimmobilized assays with a shelf life of at least 3 months. Multianalyte measurements in a complex medium are demonstrated by the simultaneous detection of both antibiotics in spiked human plasma within a sample-to-result time of less than 15 min.

[1]  J. Hacia.  Resequencing and mutational analysis using oligonucleotide microarrays , 1999, Nature Genetics.

[2]  Takehiko Kitamori,et al.  Microchip‐based immunoassay system with branching multichannels for simultaneous determination of interferon‐γ , 2002, Electrophoresis.

[3]  Guodong Liu,et al.  Electrochemical coding technology for simultaneous detection of multiple DNA targets. , 2003, Journal of the American Chemical Society.

[4]  M. Fussenegger,et al.  Broad-spectrum protein biosensors for class-specific detection of antibiotics. , 2005, Biotechnology and bioengineering.

[5]  P. Andersson,et al.  Simultaneous multiple immunoassays in a compact disc-shaped microfluidic device based on centrifugal force. , 2005, Clinical chemistry.

[6]  H. B. Halsall,et al.  Microfluidic immunosensor systems. , 2005, Biosensors & bioelectronics.

[7]  Kristen L. Helton,et al.  Microfluidic Overview of Global Health Issues Microfluidic Diagnostic Technologies for Global Public Health , 2006 .

[8]  Joseph Wang,et al.  Electrochemical biosensors: towards point-of-care cancer diagnostics. , 2006, Biosensors & bioelectronics.

[9]  G. Whitesides.  The origins and the future of microfluidics , 2006, Nature.

[10]  S. Dunbar.  Applications of Luminex® xMAP™ technology for rapid, high-throughput multiplexed nucleic acid detection , 2005, Clinica Chimica Acta.

[11]  Sylvia Kwakye,et al.  Electrochemical microfluidic biosensor for nucleic acid detection with integrated minipotentiostat. , 2006, Biosensors & bioelectronics.

[12]  Jim Kling.  Moving diagnostics from the bench to the bedside , 2006, Nature Biotechnology.

[13]  Michael S Wilson,et al.  Electrochemical multianalyte immunoassays using an array-based sensor. , 2006, Analytical chemistry.

[14]  K. Poole.  Efflux pumps as antimicrobial resistance mechanisms , 2007, Annals of medicine.

[15]  M. Fussenegger,et al.  A novel generic dipstick-based technology for rapid and precise detection of tetracycline, streptogramin and macrolide antibiotics in food samples. , 2007, Journal of biotechnology.

[16]  Kia Peyvan,et al.  CombiMatrix oligonucleotide arrays: genotyping and gene expression assays employing electrochemical detection. , 2007, Biosensors & bioelectronics.

[17]  S. Hwang,et al.  Microchip‐based multiplex electro‐immunosensing system for the detection of cancer biomarkers , 2008, Electrophoresis.

[18]  Orawon Chailapakul,et al.  Electrochemical detection for paper-based microfluidics. , 2009, Analytical chemistry.

[19]  Reinhard Niessner,et al.  A regenerable immunochip for the rapid determination of 13 different antibiotics in raw milk. , 2009, The Analyst.

[20]  H. Nikaido.  Multidrug resistance in bacteria. , 2009, Annual review of biochemistry.

[21]  Ángel Maquieira,et al.  Fast screening methods to detect antibiotic residues in food samples , 2010 .

[22]  Y. K. Cheung,et al.  1 Supplementary Information for : Microfluidics-based diagnostics of infectious diseases in the developing world , 2011 .

[23]  Seokheun Choi,et al.  Microfluidic-based biosensors toward point-of-care detection of nucleic acids and proteins , 2010, Microfluidics and nanofluidics.

[24]  Samuel K Sia,et al.  Commercialization of microfluidic point-of-care diagnostic devices. , 2012, Lab on a chip.

[25]  Kangsun Lee,et al.  Design of pressure-driven microfluidic networks using electric circuit analogy. , 2012, Lab on a chip.

[26]  James F Rusling,et al.  Multiplexed electrochemical protein detection and translation to personalized cancer diagnostics. , 2013, Analytical chemistry.

[27]  Andreas Menzel,et al.  Detection of real-time dynamics of drug-target interactions by ultralong nanowalls. , 2013, Lab on a chip.

[28]  Amy E Herr,et al.  Microfluidic Multiplexing in Bioanalyses , 2013, Journal of laboratory automation.

[29]  César Fernández-Sánchez,et al.  Diagnostics Using Multiplexed Electrochemical Readout Devices , 2014 .

[30]  Günter Gauglitz,et al.  Point-of-care platforms. , 2014, Annual review of analytical chemistry.

[31]  Mauro Ferrari,et al.  Point-of-care technologies for molecular diagnostics using a drop of blood. , 2014, Trends in biotechnology.

[32]  G. Urban,et al.  Sensitive, rapid and quantitative detection of substance P in serum samples using an integrated microfluidic immunochip. , 2014, Biosensors & bioelectronics.

[33]  Kim E. Sapsford,et al.  Evaluation of Optical Detection Platforms for Multiplexed Detection of Proteins and the Need for Point-of-Care Biosensors for Clinical Use , 2014, Sensors.

[34]  B. Fritsch,et al.  Polymer-based, flexible glutamate and lactate microsensors for in vivo applications. , 2014, Biosensors & bioelectronics.

[35]  Andreas Plückthun,et al.  Reproducibility: Standardize antibodies used in research , 2015, Nature.

[36]  G. Urban,et al.  Self-assembled magnetic bead chains for sensitivity enhancement of microfluidic electrochemical biosensor platforms. , 2015, Lab on a chip.

[37]  James F Rusling,et al.  Automated multiplexed ECL Immunoarrays for cancer biomarker proteins. , 2015, Analytical chemistry.

[38]  Rene S Schloss,et al.  Development and validation of a microfluidic immunoassay capable of multiplexing parallel samples in microliter volumes. , 2015, Lab on a chip.

[39]  A. Berlina,et al.  'Traffic light' immunochromatographic test based on multicolor quantum dots for the simultaneous detection of several antibiotics in milk. , 2015, Biosensors & bioelectronics.

[40]  G. Urban,et al.  Electrochemical Microfluidic Platform for Simultaneous Multi-analyte Detection☆ , 2015 .

[41]  Craig Watson,et al.  A Digital-Analog Microfluidic Platform for Patient-Centric Multiplexed Biomarker Diagnostics of Ultralow Volume Samples. , 2016, ACS nano.