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Emerging Loop-mediated Isothermal Amplification-based Microchip and Microdevice Technologies for Nucleic Acid Detection Mohammadali Safavieh, Manoj Kumar Kanakasabapathy, Farhang Tarlan, Minhaz uddin Ahmed, Mohammed Zourob, Waseem Asghar, and Hadi Shafiee ACS Biomater. Sci. Eng., Just Accepted Manuscript • DOI: 10.1021/acsbiomaterials.5b00449 • Publication Date (Web): 21 Jan 2016 Downloaded from http://pubs.acs.org on January 26, 2016
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ACS Biomaterials Science & Engineering
Emerging Loop-mediated Isothermal Amplificationbased Microchip and Microdevice Technologies for Nucleic Acid Detection Mohammadali Safavieh1, Manoj K. Kanakasabapathy1, Farhang Tarlan1, Minhaz U. Ahmed2, Mohammed Zourob3, Waseem Asghar4*, Hadi Shafiee1* 1
Division of Biomedical Engineering, Division of Renal Medicine, Department of Medicine,
Brigham and Women’s Hospital, Harvard Medical School, 65 Lansdowne St, Cambridge, MA 02139, USA 2
Biosensors and Biotechnology Laboratory, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong, Gadong, Negara Brunei Darussalam
3
Institute for Global Food Security, School of Biological sciences, Queen’s University Belfast, Stranmillis Road, Belfast BT9 5AG, UK.
4
Department of Computer Engineering & Electrical Engineering and Computer Science, Florida Atlantic University, 777 Glades Road, Boca Raton, FL 33431, USA
*Corresponding Authors:
[email protected], Email:
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ABSTRACT
Rapid, sensitive, and selective pathogen detection is of paramount importance in infectious disease diagnosis and monitoring of treatment. Currently available diagnostic assays based on polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) are timeconsuming, complex, and relatively expensive, thus limiting their utility in resource-limited settings. Loop-mediated isothermal amplification (LAMP) techniques have been used extensively in the development of rapid and sensitive diagnostic assays for pathogen detection and nucleic acid analysis and hold great promise for revolutionizing point-of-care molecular diagnostics. Here, we review novel LAMP-based lab-on-a-chip (LOC) diagnostic assays developed for pathogen detection over the past several years. We review various LOC platforms based on their design strategies for pathogen detection and discuss LAMP-based platforms still in development and already in the commercial pipeline. This review is intended as a guide to the use of LAMP techniques in LOC platforms for molecular diagnostics and genomic amplifications.
KEYWORDS: Nucleic acid tests, Molecular Diagnostics, Lab on a Chip, Microfluidics, LAMP, Pathogen Detection
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ACS Biomaterials Science & Engineering
1. Introduction The detection and identification of nucleic acids (NA) using point-of-care (POC) micro total analysis systems (µTAS) is a rapidly growing field with numerous clinical and industrial applications1. µTAS-based platforms enhance nucleic acid amplification by increasing the ratio of surface area to volume, improving the transfer of heat in portable lab-on-a-chip (LOC) devices for diagnostic applications.2. Nucleic acid-based POC diagnostic platforms require cell lysis, NA extraction, and purification prior to amplification3, which is of paramount importance in detecting small quantities of NA in fingerprick volumes of biological samples. Though polymerase chain reaction (PCR) is the most common method for DNA amplification, it involves multiple thermo-cycling steps. Isothermal amplification of NA eliminates the thermo-cycling steps, reducing cost and increasing assay quality4. In addition to loop-mediated isothermal amplification (LAMP), other isothermal amplification techniques, including rolling circle amplification (RCA)5, strand displacement amplification (SDA)6, signal-mediated amplification of RNA technology (SMART)7, nucleic acid sequencebased amplification (NASBA)8, single primer-triggered isothermal amplification9, helicasedependent amplification (HDA)10, and cross-priming amplification (CPA)11,12 are used to isothermally amplify NA. However, LAMP13 has been shown to be faster and more stable, sensitive, and specific for NA detection. Because of the nature of the reaction, LAMP-based methods produce >50-fold more amplicon than PCR-based techniques14. LAMP also has the ability to amplify medium- to long-range template strands of nucleic acids (>130 bp and
