Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy
Authorized Users Only
2013
Authors
Rakićević, LjiljanaKušić-Tišma, Jelena
Kovač, Mirjana
Backović, Dragana
Radojković, Dragica
Article (Published version)
Metadata
Show full item recordAbstract
During the last decade genetic factors affecting coumarin therapy have been extensively investigated. The most important genes appear to be CYP2C9 and VKORC1, and different studies have shown that DNA testing can dramatically improve the safety and effectiveness of the therapy. However, the implementation of pharmacogenetic testing in everyday practice is still not a reality. Facilities and ability to get results before the start of therapy are very important. The implementation of specific methodology and equipment for particular type of diagnostics can represent a serious, even impossible, financial hurdle to overcome (especially in developing countries). For this reason, the use of every tool that contributes to rationalization of the existing methods can be a considerable asset. Therefore, we set the goal to rationalize our current DNA sequencing based protocol for analysis of the VKORC1 c.-1639G gt A, CYP2C9*2 and CYP2C9*3 variant alleles, in order to obtain shorter and easier pro...cedure. Simplification of the protocol was achieved by setting up multiplex PCR and omitting DNA extraction. This rationalization of the existing DNA sequencing based procedure allows getting results in 12 hours. The new protocol was tested on 118 samples. Obtained results have shown full accordance to those obtained with previous, non-modified protocol. Therefore, given the circumstances, we consider that protocol for pharmocogenetic testing should be made more accessible - both to doctors and patients. It is one of the prerequisites in order to make genotyping prior to the therapy common practice.
Keywords:
VKORC1 / pharmacogenetics / CYP2C9 / coumarin therapy / coumarin derivates / AnticoagulationSource:
Scandinavian Journal of Clinical & Laboratory Investigation, 2013, 73, 6, 523-527Publisher:
- Taylor & Francis Ltd, Abingdon
Funding / projects:
- Complex diseases as a model system for phenotype modulation- structural and functional analysis of molecular biomarkers (RS-MESTD-Basic Research (BR or ON)-173008)
DOI: 10.3109/00365513.2013.809142
ISSN: 0036-5513
PubMed: 23806093
WoS: 000324400500009
Scopus: 2-s2.0-84884234408
Collections
Institution/Community
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Rakićević, Ljiljana AU - Kušić-Tišma, Jelena AU - Kovač, Mirjana AU - Backović, Dragana AU - Radojković, Dragica PY - 2013 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/651 AB - During the last decade genetic factors affecting coumarin therapy have been extensively investigated. The most important genes appear to be CYP2C9 and VKORC1, and different studies have shown that DNA testing can dramatically improve the safety and effectiveness of the therapy. However, the implementation of pharmacogenetic testing in everyday practice is still not a reality. Facilities and ability to get results before the start of therapy are very important. The implementation of specific methodology and equipment for particular type of diagnostics can represent a serious, even impossible, financial hurdle to overcome (especially in developing countries). For this reason, the use of every tool that contributes to rationalization of the existing methods can be a considerable asset. Therefore, we set the goal to rationalize our current DNA sequencing based protocol for analysis of the VKORC1 c.-1639G gt A, CYP2C9*2 and CYP2C9*3 variant alleles, in order to obtain shorter and easier procedure. Simplification of the protocol was achieved by setting up multiplex PCR and omitting DNA extraction. This rationalization of the existing DNA sequencing based procedure allows getting results in 12 hours. The new protocol was tested on 118 samples. Obtained results have shown full accordance to those obtained with previous, non-modified protocol. Therefore, given the circumstances, we consider that protocol for pharmocogenetic testing should be made more accessible - both to doctors and patients. It is one of the prerequisites in order to make genotyping prior to the therapy common practice. PB - Taylor & Francis Ltd, Abingdon T2 - Scandinavian Journal of Clinical & Laboratory Investigation T1 - Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy EP - 527 IS - 6 SP - 523 VL - 73 DO - 10.3109/00365513.2013.809142 ER -
@article{ author = "Rakićević, Ljiljana and Kušić-Tišma, Jelena and Kovač, Mirjana and Backović, Dragana and Radojković, Dragica ", year = "2013", abstract = "During the last decade genetic factors affecting coumarin therapy have been extensively investigated. The most important genes appear to be CYP2C9 and VKORC1, and different studies have shown that DNA testing can dramatically improve the safety and effectiveness of the therapy. However, the implementation of pharmacogenetic testing in everyday practice is still not a reality. Facilities and ability to get results before the start of therapy are very important. The implementation of specific methodology and equipment for particular type of diagnostics can represent a serious, even impossible, financial hurdle to overcome (especially in developing countries). For this reason, the use of every tool that contributes to rationalization of the existing methods can be a considerable asset. Therefore, we set the goal to rationalize our current DNA sequencing based protocol for analysis of the VKORC1 c.-1639G gt A, CYP2C9*2 and CYP2C9*3 variant alleles, in order to obtain shorter and easier procedure. Simplification of the protocol was achieved by setting up multiplex PCR and omitting DNA extraction. This rationalization of the existing DNA sequencing based procedure allows getting results in 12 hours. The new protocol was tested on 118 samples. Obtained results have shown full accordance to those obtained with previous, non-modified protocol. Therefore, given the circumstances, we consider that protocol for pharmocogenetic testing should be made more accessible - both to doctors and patients. It is one of the prerequisites in order to make genotyping prior to the therapy common practice.", publisher = "Taylor & Francis Ltd, Abingdon", journal = "Scandinavian Journal of Clinical & Laboratory Investigation", title = "Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy", pages = "527-523", number = "6", volume = "73", doi = "10.3109/00365513.2013.809142" }
Rakićević, L., Kušić-Tišma, J., Kovač, M., Backović, D.,& Radojković, D.. (2013). Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy. in Scandinavian Journal of Clinical & Laboratory Investigation Taylor & Francis Ltd, Abingdon., 73(6), 523-527. https://doi.org/10.3109/00365513.2013.809142
Rakićević L, Kušić-Tišma J, Kovač M, Backović D, Radojković D. Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy. in Scandinavian Journal of Clinical & Laboratory Investigation. 2013;73(6):523-527. doi:10.3109/00365513.2013.809142 .
Rakićević, Ljiljana, Kušić-Tišma, Jelena, Kovač, Mirjana, Backović, Dragana, Radojković, Dragica , "Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy" in Scandinavian Journal of Clinical & Laboratory Investigation, 73, no. 6 (2013):523-527, https://doi.org/10.3109/00365513.2013.809142 . .