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The sweet detection...
The sweet detection of rolling circle amplification : Glucose-based electrochemical genosensor for the detection of viral nucleic acid
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- Ciftci, S. (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)
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- Cánovas, Rocio (författare)
- KTH,Tillämpad fysikalisk kemi
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- Neumann, Felix (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)
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Paulraj, T. (författare)
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- Nilsson, Mats (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)
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- Crespo, Gaston A., 1980- (författare)
- KTH,Tillämpad fysikalisk kemi
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- Madaboosi, Narayanan (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)
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(creator_code:org_t)
- Elsevier Ltd, 2020
- 2020
- Engelska.
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Ingår i: Biosensors & bioelectronics. - : Elsevier Ltd. - 0956-5663 .- 1873-4235. ; 151
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Herein, an isothermal padlock probe-based assay for the simple and portable detection of pathogens coupled with a glucose oxidase (GOx)-based electrochemical readout is reported. Infectious diseases remain a constant threat on a global scale, as in recurring pandemics. Rapid and portable diagnostics hold the promise to tackle the spreading of diseases and decentralising healthcare to point-of-care needs. Ebola, a hypervariable RNA virus causing fatalities of up to 90% for recent outbreaks in Africa, demands immediate attention for bedside diagnostics. The design of the demonstrated assay consists of a rolling circle amplification (RCA) technique, responsible for the generation of nucleic acid amplicons as RCA products (RCPs). The RCPs are generated on magnetic beads (MB) and subsequently, connected via streptavidin-biotin bonds to GOx. The enzymatic catalysis of glucose by the bound GOx allows for an indirect electrochemical measurement of the DNA target. The RCPs generated on the surface of the MB were confirmed by scanning electron microscopy, and among other experimental conditions such as the type of buffer, temperature, concentration of GOx, sampling and measurement time were evaluated for the optimum electrochemical detection. Accordingly, 125 μg mL−1 of GOx with 5 mM glucose using phosphate buffer saline (PBS), monitored for 1 min were selected as the ideal conditions. Finally, we assessed the analytical performance of the biosensing strategy by using clinical samples of Ebola virus from patients. Overall, this work provides a proof-of-concept bioassay for simple and portable molecular diagnostics of emerging pathogens using electrochemical detection, especially in resource-limited settings.
Ämnesord
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
Nyckelord
- Chronoamperometric readout
- Ebola virus
- Glucose oxidase
- Molecular diagnostics
- Point-of-care
- Rolling-circle-amplification (RCA)
- Diagnosis
- Diseases
- Glucose
- Glucose sensors
- Nucleic acids
- Product design
- Scanning electron microscopy
- Viruses
- Point of care
- Rolling circle amplifications
- Chemical detection
- biotin
- buffer
- phosphate buffered saline
- streptavidin
- virus DNA
- Africa
- amplicon
- analytical parameters
- Article
- bioassay
- biological monitoring
- catalysis
- concentration (parameter)
- controlled study
- Ebola hemorrhagic fever
- Ebolavirus
- electrochemical detection
- enzyme binding
- epidemic
- gene targeting
- isotherm
- magnetism
- measurement
- molecular probe
- nonhuman
- nucleic acid amplification
- pandemic
- point of care testing
- rolling circle amplification
- sampling
- surface property
- temperature sensitivity
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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