title: High-throughput real-time electrochemical monitoring of LAMP for pathogenic bacteria detection.
abstract: One of the significant challenges in healthcare is the development of point-of-care (POC) diagnostics. POC diagnostics require low-cost devices that offer portability, simplicity in operation and the ability for high-throughput and quantitative analysis. Here, we present a novel roll-to-roll ribbon fluid-handling device for electrochemical real-time monitoring of nucleic acid (NA) amplification and bacteria detection. The device rendered loop-mediated isothermal amplification (LAMP) and real-time electrochemical detection based on the interaction between LAMP amplicon and the redox-reactive osmium complex. We have shown the detection of 30CFU/ml of Escherichia coli (in the range between 30 and 3×10(7)CFU/ml) and 200CFU/ml of Staphylococcus aureus (in the range of 200-2×10(5)CFU/ml) cultured samples in both real-time and end point detection. This device can be used for the detection of various Gram-negative and a number of Gram-positive bacterial pathogens with high sensitivity and specificity in a high-throughput format. Using a roll-to-roll cassette approach, we could detect 12 samples in one assay. Since the LAMP and electrochemical analysis are implemented within sealed flexible biochips, time-consuming processing steps are not required and the risk of contamination is significantly reduced.
authors.affiliation: Institut national de la recherche scientifique, Énergie, Matériaux et Télécommunications (INRS-EMT), Université du Québec, 1650 Boulevard Lionel-Boulet, Varennes (Québec) J3X1S2, Canada.
authors.forename: Mohammadali
authors.initials: M
authors.lastname: Safavieh
authors.affiliation: Institut national de la recherche scientifique, Énergie, Matériaux et Télécommunications (INRS-EMT), Université du Québec, 1650 Boulevard Lionel-Boulet, Varennes (Québec) J3X1S2, Canada; Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong BE 1410, Brunei Darussalam.
authors.forename: Minhaz Uddin
authors.initials: MU
authors.lastname: Ahmed
authors.affiliation: Center of Biomedical Engineering, Vincent Building, Cranfield University, Bedfordshire MK43 0AL, United Kingdom.
authors.forename: Andy
authors.initials: A
authors.lastname: Ng
authors.affiliation: Center of Biomedical Engineering, Vincent Building, Cranfield University, Bedfordshire MK43 0AL, United Kingdom. Electronic address: m.zourob@cranfield.ac.uk.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Zourob
mesh_terms: D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification*; D058491:Bacterial Load / Q000295:instrumentation*; D003199:Computer Systems; D003217:Conductometry / Q000295:instrumentation*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D057166:High-Throughput Screening Assays / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation* / Q000379:methods
chemical_list: D004269:DNA, Bacterial
keywords: Bacteria detection; Cassette POC; LAMP; Point of care device; Roll to roll