Publication: In-situ monitoring of real-time loop-mediated isothermal amplification with qcm: Detecting listeria monocytogenes
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Issued Date
2021
Resource Type
Language
eng
File Type
application/pdf
ISSN
20796374
Other identifier(s)
2-s2.0-85114631824
Rights Holder(s)
Scopus
Bibliographic Citation
Biosensors. Vol 11, No.9 (2021)
Suggested Citation
Wachiralurpan S., Phung-On I., Chanlek N., Areekit S., Chansiri K., Lieberzeit P.A. In-situ monitoring of real-time loop-mediated isothermal amplification with qcm: Detecting listeria monocytogenes. Biosensors. Vol 11, No.9 (2021). doi:10.3390/bios11090308 Retrieved from: https://hdl.handle.net/20.500.14740/4089
Abstract
Functionalized DNA sequences are promising sensing elements to combine with transducers for bio-sensing specific target microbes. As an application example, this paper demonstrates in situ detection of loop-mediated isothermal amplification products by hybridizing them with thiolated-ssDNA covalently anchored on the electrodes of a quartz crystal microbalance (QCM). Such hybridization leads to a frequency signal, which is suitable for monitoring real-time LAMP amplification based on mass-sensing: it detects interactions between the complementary nucleobases of LAMP products in solution and the thiolated-ssDNA probe sequence on the gold surface. Target DNA LAMP products cause irreversible frequency shifts on the QCM surfaces during hybridization in the kHz range, which result from both changes in mass and charge on the electrode surface. In order to confirm the LAMP assay working in the QCM sensing system at elevated temperature, the sky blue of positive LAMP products solution was achieved by using the Hydroxy Naphthol Blue (HNB) and agarose gel electrophoresis. Since on-QCM sensing of DNA hybridization leads to irreversible sensor responses, this work shows characterization by X-ray photoelectron spectroscopy (XPS) core spectra of S2p, N1s, Mg1s, P2p and C1s. XPS results confirmed that indeed both DNA and by-products of LAMP attached to the surface. Listeria monocytogenes DNA served to study in-situ detection of amplified LAMP products on DNA-functionalized surfaces. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Subject(s)
Complement component C1
Cysteine
Gold
Hydroxy naphthol blue
Naphthalene derivative
Nucleic acid base
Single stranded DNA
Unclassified drug
Agar gel electrophoresis
Article
Bacterium detection
Chemical reaction
Controlled study
Covalent bond
DNA hybridization
Listeria monocytogenes
Loop mediated isothermal amplification
Monitoring
Nonhuman
Quartz crystal microbalance
Temperature
X ray photoemission spectroscopy
Genetics
Molecular diagnosis
Nucleic acid amplification
Quartz crystal microbalance
Listeria monocytogenes
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques
Quartz Crystal Microbalance Techniques
Cysteine
Gold
Hydroxy naphthol blue
Naphthalene derivative
Nucleic acid base
Single stranded DNA
Unclassified drug
Agar gel electrophoresis
Article
Bacterium detection
Chemical reaction
Controlled study
Covalent bond
DNA hybridization
Listeria monocytogenes
Loop mediated isothermal amplification
Monitoring
Nonhuman
Quartz crystal microbalance
Temperature
X ray photoemission spectroscopy
Genetics
Molecular diagnosis
Nucleic acid amplification
Quartz crystal microbalance
Listeria monocytogenes
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques
Quartz Crystal Microbalance Techniques