Visual Detection of Stem-Loop Primer Amplification (SPA) Products without Denaturation Using Peroxidase-like DNA Machines (PxDM)

Rapid, inexpensive, and accurate determination of nucleic acids is a decisive factor in evaluating population's health and monitoring treatment at point-of-care (POC) settings. Testing systems with visual outputs can provide instrument-free signal detection. Isothermal amplification technologie...

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Bibliographic Details
Published in:International journal of molecular sciences 2023-04, Vol.24 (9), p.7812
Main Authors: Maltzeva, Yulia I, Gorbenko, Daria A, Nikitina, Ekaterina V, Rubel, Maria S, Kolpashchikov, Dmitry M
Format: Article
Language:English
Subjects:
Denaturation
Deoxyribonucleic acid
Design
DNA
E coli
Epstein-Barr virus
Gene amplification
Genetic testing
Nucleic Acid Amplification Techniques - methods
Nucleic Acids
Nucleotides
Pathogens
Peroxidase
Peroxidases
Polymerase chain reaction
Polymerase Chain Reaction - methods
Sensitivity and Specificity
Sensors
Signal detection
Substitution reactions
Visual signals
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Summary:Rapid, inexpensive, and accurate determination of nucleic acids is a decisive factor in evaluating population's health and monitoring treatment at point-of-care (POC) settings. Testing systems with visual outputs can provide instrument-free signal detection. Isothermal amplification technologies can substitute conventional polymerase chain reaction (PCR) testing due to compatibility with the POC diagnostic. Here, we have visually detected DNA fragments obtained by stem-loop-primer-assisted isothermal amplification (SPA), but not those obtained by PCR or LAMP amplification using DNA nanomachines with peroxidase-like activity (PxDM) with sensitivity to a single nucleotide substitution. Compared to the diagnostics with conventional loop-mediated isothermal amplification (LAMP), the PxDM method produces no false positive signals with the non-specific amplification products. The study suggests that PxDM, in conjunction with SPA isothermal amplification, can become a valid platform for POC testing systems.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms24097812