Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR
Samo za registrovane korisnike
2013
Autori
Jovanović, Katarina KSavić, Aleksandar G
Janković, Radmila N
Radulović, Sinisa S
Spasić, Slađana
Radotić, Ksenija
Članak u časopisu (Objavljena verzija)
Metapodaci
Prikaz svih podataka o dokumentuApstrakt
The key method for therapies of various cancer types could be the molecular-targeted therapy, based on individual gene profile for each patient. One of the main procedures used for genetic testing is the real-time polymerase chain reaction (real-time PCR). Physical principle behind real-time PCR procedure is the fluorescence. Fluorescence labeled probes (primers) is attached to quenchers. Upon reaction of polymerization, quenchers are removed, and the fluorescence emission intensity increases in time. Emission spectra shape and its maximum position can differ if the fluorophore was present in different microenvironment. That property is widely exploited in fluorescence spectroscopy and chromatography. This paper, for the first time, describes utilization of full spectroscopic potential of multichannel excitation/emission filter sets in real-time PCR device. Instead of monitoring fluorescence intensity in time for a single fluorescence emission channel, the ratio values of three differe...nt kinetics curves were calculated and analyzed by applying k-means clustering and dendrogram analysis. Obtained results have shown that described analytical improvement provides identification of nine different groups of mutations if the commercial QIAGEN (R) EGFR PCR Kit was used. Method can be applied to any kit, capable to simultaneously detect several different mutations.
Ključne reči:
Detection of DNA mutations / excitation fluorescence / emission fluorescence / real-time PCRIzvor:
Medical Hypotheses, 2013, 80, 4, 376-379Izdavač:
- Churchill Livingstone, Edinburgh
Finansiranje / projekti:
- Ispitivanja odnosa struktura-funkcija u ćelijskom zidu biljaka i izmene strukture zida enzimskim inženjeringom (RS-MESTD-Basic Research (BR or ON)-173017)
- Ribe kao bioindikatori stanja kvaliteta otvorenih voda Srbije (RS-MESTD-Basic Research (BR or ON)-173045)
- Farmakodinamska i farmakogenomska ispitivanja novijih lekova u lečenju solidnih tumora (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41026)
DOI: 10.1016/j.mehy.2013.01.004
ISSN: 0306-9877
PubMed: 23352288
WoS: 000317164500009
Scopus: 2-s2.0-84875404058
Institucija/grupa
Institut za multidisciplinarna istraživanjaTY - JOUR AU - Jovanović, Katarina K AU - Savić, Aleksandar G AU - Janković, Radmila N AU - Radulović, Sinisa S AU - Spasić, Slađana AU - Radotić, Ksenija PY - 2013 UR - http://rimsi.imsi.bg.ac.rs/handle/123456789/702 AB - The key method for therapies of various cancer types could be the molecular-targeted therapy, based on individual gene profile for each patient. One of the main procedures used for genetic testing is the real-time polymerase chain reaction (real-time PCR). Physical principle behind real-time PCR procedure is the fluorescence. Fluorescence labeled probes (primers) is attached to quenchers. Upon reaction of polymerization, quenchers are removed, and the fluorescence emission intensity increases in time. Emission spectra shape and its maximum position can differ if the fluorophore was present in different microenvironment. That property is widely exploited in fluorescence spectroscopy and chromatography. This paper, for the first time, describes utilization of full spectroscopic potential of multichannel excitation/emission filter sets in real-time PCR device. Instead of monitoring fluorescence intensity in time for a single fluorescence emission channel, the ratio values of three different kinetics curves were calculated and analyzed by applying k-means clustering and dendrogram analysis. Obtained results have shown that described analytical improvement provides identification of nine different groups of mutations if the commercial QIAGEN (R) EGFR PCR Kit was used. Method can be applied to any kit, capable to simultaneously detect several different mutations. PB - Churchill Livingstone, Edinburgh T2 - Medical Hypotheses T1 - Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR EP - 379 IS - 4 SP - 376 VL - 80 DO - 10.1016/j.mehy.2013.01.004 ER -
@article{ author = "Jovanović, Katarina K and Savić, Aleksandar G and Janković, Radmila N and Radulović, Sinisa S and Spasić, Slađana and Radotić, Ksenija", year = "2013", abstract = "The key method for therapies of various cancer types could be the molecular-targeted therapy, based on individual gene profile for each patient. One of the main procedures used for genetic testing is the real-time polymerase chain reaction (real-time PCR). Physical principle behind real-time PCR procedure is the fluorescence. Fluorescence labeled probes (primers) is attached to quenchers. Upon reaction of polymerization, quenchers are removed, and the fluorescence emission intensity increases in time. Emission spectra shape and its maximum position can differ if the fluorophore was present in different microenvironment. That property is widely exploited in fluorescence spectroscopy and chromatography. This paper, for the first time, describes utilization of full spectroscopic potential of multichannel excitation/emission filter sets in real-time PCR device. Instead of monitoring fluorescence intensity in time for a single fluorescence emission channel, the ratio values of three different kinetics curves were calculated and analyzed by applying k-means clustering and dendrogram analysis. Obtained results have shown that described analytical improvement provides identification of nine different groups of mutations if the commercial QIAGEN (R) EGFR PCR Kit was used. Method can be applied to any kit, capable to simultaneously detect several different mutations.", publisher = "Churchill Livingstone, Edinburgh", journal = "Medical Hypotheses", title = "Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR", pages = "379-376", number = "4", volume = "80", doi = "10.1016/j.mehy.2013.01.004" }
Jovanović, K. K., Savić, A. G., Janković, R. N., Radulović, S. S., Spasić, S.,& Radotić, K.. (2013). Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR. in Medical Hypotheses Churchill Livingstone, Edinburgh., 80(4), 376-379. https://doi.org/10.1016/j.mehy.2013.01.004
Jovanović KK, Savić AG, Janković RN, Radulović SS, Spasić S, Radotić K. Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR. in Medical Hypotheses. 2013;80(4):376-379. doi:10.1016/j.mehy.2013.01.004 .
Jovanović, Katarina K, Savić, Aleksandar G, Janković, Radmila N, Radulović, Sinisa S, Spasić, Slađana, Radotić, Ksenija, "Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR" in Medical Hypotheses, 80, no. 4 (2013):376-379, https://doi.org/10.1016/j.mehy.2013.01.004 . .