title: Low-cost, real-time, continuous flow PCR system for pathogen detection.
abstract: In this paper, we present a portable and low cost point-of-care (POC) PCR system for quantitative detection of pathogens. Our system is based on continuous flow PCR which maintains fixed temperatures zones and pushes the PCR solution between two heated areas allowing for faster heat transfer and as a result, a faster PCR. The PCR system is built around a 46.0 mm × 30.9 mm × 0.4 mm disposable thermoplastic chip. In order to make the single-use chip economically viable, it was manufactured by hot embossing and was designed to be compatible with roll-to-roll embossing for large scale production. The prototype instrumentation surrounding the chip includes two heaters, thermal sensors, and an optical system. The optical system allows for pathogen detection via real time fluorescence measurements. FAM probes were used as fluorescent reporters of the amplicons generated during the PCR. To demonstrate the function of the chip, two infectious bacteria targets were selected: Chlamydia trachomatis and Escherichia coli O157:H7. For both bacteria, the limit of detection of the system was determined, PCR efficiencies were calculated, and different flow velocities were tested. We have demonstrated successful detection for these two bacterial pathogens highlighting the versatility and broad utility of our portable, low-cost, and rapid PCR diagnostic device.
authors.affiliation: Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA.|Grup d'Enginyeria de Materials (GEMAT), Institut Químic de Sarrià, Universitat Ramón Lllull, Barcelona, Spain.
authors.forename: B Leticia
authors.initials: BL
authors.lastname: Fernández-Carballo
authors.affiliation: Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA.
authors.forename: Ian
authors.initials: I
authors.lastname: McGuiness
authors.affiliation: Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: McBeth
authors.affiliation: Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA.
authors.forename: Maxim
authors.initials: M
authors.lastname: Kalashnikov
authors.affiliation: Grup d'Enginyeria de Materials (GEMAT), Institut Químic de Sarrià, Universitat Ramón Lllull, Barcelona, Spain.
authors.forename: Salvador
authors.initials: S
authors.lastname: Borrós
authors.affiliation: Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA.|Mechanical Engineering Department, Boston University, Boston, MA, USA.
authors.forename: Andre
authors.initials: A
authors.lastname: Sharon
authors.affiliation: Fraunhofer Center for Manufacturing Innovation, Brookline, MA, USA. asauerbudge@fraunhofer.org.|Biomedical Engineering Department, Boston University, Boston, MA, USA. asauerbudge@fraunhofer.org.
authors.forename: Alexis F
authors.identifier: http://orcid.org/0000-0001-5961-5286
authors.initials: AF
authors.lastname: Sauer-Budge
mesh_terms: D002692:Chlamydia trachomatis / Q000235:genetics* / Q000302:isolation & purification*; D003365:Costs and Cost Analysis; D004867:Equipment Design; D019453:Escherichia coli O157 / Q000235:genetics* / Q000302:isolation & purification*; D046208:Microchip Analytical Procedures; D019095:Point-of-Care Systems; D060888:Real-Time Polymerase Chain Reaction / Q000191:economics* / Q000295:instrumentation*
keywords: Lab-on-a-chip; Nucleic acid test; On chip polymerase chain reaction (PCR); Pathogen diagnostic; Point-of-care (POC); Real-time polymerase chain reaction (qPCR)