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 3×107CFU/ml) and 200CFU/ml of Staphylococcus aureus (in the range of 200-2×105CFU/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. © 2014 Elsevier B.V.