title: A novel, sensitive and label-free loop-mediated isothermal amplification detection method for nucleic acids using luminophore dyes.
abstract: Electrochemiluminescence (ECL) has been widely rendered for nucleic acid testing. Here, we integrate loop-mediated isothermal amplification (LAMP) with ECL technique for DNA detection and quantification. The target LAMP DNA bound electrostatically with [Ru(bpy)3](+2) on the carbon electrode surface, and an ECL reaction was triggered by tripropylamine (TPrA) to yield luminescence. We illustrated this method as a new and highly sensitive strategy for the detection of sequence-specific DNA from different meat species at picogram levels. The proposed strategy renders the signal amplification capacities of TPrA and combines LAMP with inherently high sensitivity of the ECL technique, to facilitate the detection of low quantities of DNA. By leveraging this technique, target DNA of Sus scrofa (pork) meat was detected as low as 1pg/µL (3.43×10(-1)copies/µL). In addition, the proposed technique was applied for detection of Bacillus subtilis DNA samples and detection limit of 10pg/µL (2.2×10(3)copies/µL) was achieved. The advantages of being isothermal, sensitive and robust with ability for multiplex detection of bio-analytes makes this method a facile and appealing sensing modality in hand-held devices to be used at the point-of-care (POC).
authors.affiliation: Biosensors and Biotechnology Laboratory, Chemical Science Programme, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong, BE 1410 Brunei Darussalam.
authors.forename: Sharmili
authors.initials: S
authors.lastname: Roy
authors.affiliation: Biosensors and Biotechnology Laboratory, Chemical Science Programme, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong, BE 1410 Brunei Darussalam.
authors.forename: Sim Xiao
authors.initials: SX
authors.lastname: Wei
authors.affiliation: Biosensors and Biotechnology Laboratory, Chemical Science Programme, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong, BE 1410 Brunei Darussalam.
authors.forename: Jean Liew Zhi
authors.initials: JLZ
authors.lastname: Ying
authors.affiliation: Division of Biomedical Engineering, Division of Renal Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: Mohammadali
authors.initials: M
authors.lastname: Safavieh
authors.affiliation: Biosensors and Biotechnology Laboratory, Chemical Science Programme, Faculty of Science, Universiti Brunei Darussalam, Jalan Tungku Link, Gadong, BE 1410 Brunei Darussalam. Electronic address: minhaz.ahmed@ubd.edu.bn.
authors.forename: Minhaz Uddin
authors.initials: MU
authors.lastname: Ahmed
mesh_terms: D003217:Conductometry / Q000295:instrumentation*; D004247:DNA / Q000032:analysis / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D049408:Luminescent Agents / Q000737:chemistry*; D008163:Luminescent Measurements / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D009696:Nucleic Acids; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D013194:Staining and Labeling
chemical_list: D049408:Luminescent Agents; D009696:Nucleic Acids; D004247:DNA
keywords: Bacillus subtilis; Electrochemiluminescence; Loop-mediated isothermal amplification; Nucleic acid detection; Sus scrofa