Designing a fluorescence padlock probe-based biosensor and colorimetric assay for the detection of G12D KRAS mutation

Autor:	Mohammad Hossein Ghahremani, Gholamreza Tavoosidana, Fatemeh Mahmoudian, Mostafa Akbariqomi, Reza Heidari, Samaneh Fathi, Nader Roshan, Fatemeh Karimi, Samira Bahrami, Moloud Absalan, Mahdi Adabi
Rok vydání:	2021
Předmět:	Detection limit Chromatography business.industry Point mutation Biochemistry (medical) Clinical Biochemistry medicine.disease_cause Target concentration Fluorescence Rolling circle replication Drug Discovery medicine KRAS business Biosensor Kras mutation
Zdroj:	Biomarkers in Medicine. 15:1741-1754
ISSN:	1752-0371 1752-0363
Popis:	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°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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::c5a9fe8f9195728be81747da8c6ec92a https://doi.org/10.2217/bmm-2021-0333 Zobrazit plný text záznamu