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Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers

BACKGROUND: Genetic polymorphisms of drug metabolisms by cytochrome P450 (P450s) could affect drug response, attracting particular interest in the pharmacogenetics. Due to the importance of CYP2C19* 17 allele and its capability of super- fast metabolism and also lack of information about distributio...

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Autores principales: Ghasemi, Zahra, Hashemi, Mehrdad, Ejabati, Mahsa, Ebrahimi, Seyyed Meisam, Kheiri Manjili, Hamidreza, Sharafi, Ali, Ramazani, Ali
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Avicenna Research Institute 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5124257/
https://www.ncbi.nlm.nih.gov/pubmed/27920888
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author Ghasemi, Zahra
Hashemi, Mehrdad
Ejabati, Mahsa
Ebrahimi, Seyyed Meisam
Kheiri Manjili, Hamidreza
Sharafi, Ali
Ramazani, Ali
author_facet Ghasemi, Zahra
Hashemi, Mehrdad
Ejabati, Mahsa
Ebrahimi, Seyyed Meisam
Kheiri Manjili, Hamidreza
Sharafi, Ali
Ramazani, Ali
author_sort Ghasemi, Zahra
collection PubMed
description BACKGROUND: Genetic polymorphisms of drug metabolisms by cytochrome P450 (P450s) could affect drug response, attracting particular interest in the pharmacogenetics. Due to the importance of CYP2C19* 17 allele and its capability of super- fast metabolism and also lack of information about distribution of the alleles in Iranian population, this research aimed to use High Resolution Melting (HRM) method compared to PCR-RFLP for genotyping healthy Iranian population. METHODS: Blood samples were collected from 100 healthy Iranian volunteers. DNA was extracted by salting out method. Real-time PCR was used for amplification of the CYP2C19 gene and the alleles were identified by HRM. Sequencing was used to confirm the amplified DNA fragments and data were analyzed using SPSS software ver.18. RESULTS: The frequency of alleles CYP2C19*1/*1, CYP2C19*1/*17 and CYP2C19*17/*17 were estimated as 58.33, 29.1 and 11.1%, respectively. Specificity and sensitivity of HRM method were 90% and 100%, with respect to PCR-RFLP. Also, HRM analysis has been evaluated as a faster and more effective approach. CONCLUSION: Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed method is rapid, accurate, fast and economic to study the CYP2C19*17 allele and it is appropriate for other similar population genetic studies.
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spelling pubmed-51242572016-12-05 Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers Ghasemi, Zahra Hashemi, Mehrdad Ejabati, Mahsa Ebrahimi, Seyyed Meisam Kheiri Manjili, Hamidreza Sharafi, Ali Ramazani, Ali Avicenna J Med Biotechnol Original Article BACKGROUND: Genetic polymorphisms of drug metabolisms by cytochrome P450 (P450s) could affect drug response, attracting particular interest in the pharmacogenetics. Due to the importance of CYP2C19* 17 allele and its capability of super- fast metabolism and also lack of information about distribution of the alleles in Iranian population, this research aimed to use High Resolution Melting (HRM) method compared to PCR-RFLP for genotyping healthy Iranian population. METHODS: Blood samples were collected from 100 healthy Iranian volunteers. DNA was extracted by salting out method. Real-time PCR was used for amplification of the CYP2C19 gene and the alleles were identified by HRM. Sequencing was used to confirm the amplified DNA fragments and data were analyzed using SPSS software ver.18. RESULTS: The frequency of alleles CYP2C19*1/*1, CYP2C19*1/*17 and CYP2C19*17/*17 were estimated as 58.33, 29.1 and 11.1%, respectively. Specificity and sensitivity of HRM method were 90% and 100%, with respect to PCR-RFLP. Also, HRM analysis has been evaluated as a faster and more effective approach. CONCLUSION: Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed method is rapid, accurate, fast and economic to study the CYP2C19*17 allele and it is appropriate for other similar population genetic studies. Avicenna Research Institute 2016 /pmc/articles/PMC5124257/ /pubmed/27920888 Text en Copyright© 2016 Avicenna Research Institute This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.
spellingShingle Original Article
Ghasemi, Zahra
Hashemi, Mehrdad
Ejabati, Mahsa
Ebrahimi, Seyyed Meisam
Kheiri Manjili, Hamidreza
Sharafi, Ali
Ramazani, Ali
Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
title Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
title_full Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
title_fullStr Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
title_full_unstemmed Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
title_short Development of a High-Resolution Melting Analysis Method for CYP2C19*17 Genotyping in Healthy Volunteers
title_sort development of a high-resolution melting analysis method for cyp2c19*17 genotyping in healthy volunteers
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5124257/
https://www.ncbi.nlm.nih.gov/pubmed/27920888
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