期刊
BIOMARKERS IN MEDICINE
卷 15, 期 18, 页码 1741-1754出版社
FUTURE MEDICINE LTD
DOI: 10.2217/bmm-2021-0333
关键词
biosensor; colorimetric assay; HRCA; KRAS mutation; padlock probe
资金
- Tehran University of Medical Sciences [97-01-87-37598]
The study aimed to design an optimized biosensor to detect the KRAS G12D mutation in plasma using padlock probe-based hyperbranched rolling circle amplification. The results showed the biosensor has a low detection limit and linear response in detecting KRAS point mutations in liquid state reactions.
Aim: Cell-free DNA in the plasma is known to be a potential biomarker for noninvasive diagnosis of oncogenic mutations. The authors aimed to design an optimized padlock probe-based hyperbranched rolling circle amplification biosensor to detect the KRAS G12D mutation using fluorescence and colorimetric methods. Methods: Single-factor experiments, Plackett-Burman design and response surface methodology were applied to optimize the padlock probe-based hyperbranched rolling circle amplification reaction. Results: The maximum fluorescence intensity was achieved at a padlock probe concentration of 1.5 pM and target concentration of 9 pM at 38 degrees C ligation temperature. The proposed biosensor has a low detection limit of 60 fM of target DNA and a linear response in the concentration range of 60 fM to 0.2 pM. Conclusion: The results indicated the power of these assays to detect KRAS point mutations in liquid state reactions.
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