Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR
Background: A significant role of Renin-angiotensin system (RAS) genetic variants in the pathogenesis of essential hypertension and cardiovascular diseases has been proved. This study aimed to develop a new, fast and cheap method for the simultaneous detection of two missense single nucleotide polym...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2017-05-01
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| Series: | Iranian Journal of Public Health |
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| Online Access: | https://ijph.tums.ac.ir/index.php/ijph/article/view/10085 |
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| author | Mehri KHATAMI Mohammad Mehdi HEIDARI Mehdi HADADZADEH Barbara SCHEIBER-MOJDEHKAR Morteza BITARAF SANI Massoud HOUSHMAND |
| author_facet | Mehri KHATAMI Mohammad Mehdi HEIDARI Mehdi HADADZADEH Barbara SCHEIBER-MOJDEHKAR Morteza BITARAF SANI Massoud HOUSHMAND |
| author_sort | Mehri KHATAMI |
| collection | DOAJ |
| description | Background: A significant role of Renin-angiotensin system (RAS) genetic variants in the pathogenesis of essential hypertension and cardiovascular diseases has been proved. This study aimed to develop a new, fast and cheap method for the simultaneous detection of two missense single nucleotide polymorphisms (T207M or rs4762 and M268T orrs699) of angiotensinogen (AGT) in single-step Multiplex Hexa-Primer Amplification Refractory Mutation System - polymerase chain reaction (H-ARMS-PCR).
Methods: In this case-control study, 148 patients with coronary artery disease (CAD) and 135 controls were included. The patients were referred to cardiac centers in Afshar Hospital (Yazd, Iran) from 2012 to 2015. Two sets of inner primer (for each SNP) and one set outer primer pairs were designed for genotyping of rs4762 and rs699 in single tube H-ARMS-PCR. Direct sequencing of all samples was also performed to assess the accuracy of this method. DNA sequencing method validated the results of single tube H-ARMS-PCR.
Results: We found full accordance for genotype adscription by sequencing method. The frequency of the AGT T521 and C702 alleles was significantly higher in CAD patients than in the control group (OR: 0.551, 95% CI: 0.359-0.846, P=0.008 and OR: 0.629, 95% CI: 0.422-0.936, P=0.028, respectively).
Conclusion: This is the first work describing a rapid, low-cost, high-throughput simultaneous detection of rs4762 and rs699 polymorphisms in AGT gene, used in large clinical studies. |
| first_indexed | 2024-12-23T14:12:50Z |
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| issn | 2251-6085 2251-6093 |
| language | English |
| last_indexed | 2024-12-23T14:12:50Z |
| publishDate | 2017-05-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Public Health |
| spelling | doaj.art-622a72b7d4a44c81805de908c7457d242022-12-21T17:44:00ZengTehran University of Medical SciencesIranian Journal of Public Health2251-60852251-60932017-05-01466Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCRMehri KHATAMI0Mohammad Mehdi HEIDARI1Mehdi HADADZADEH2Barbara SCHEIBER-MOJDEHKAR3Morteza BITARAF SANI4Massoud HOUSHMAND5Dept. of Biology, Faculty of Sciences, Yazd University, Yazd, IranDept. of Biology, Faculty of Sciences, Yazd University, Yazd, IranDept. of Cardiac Surgery, Afshar Hospital, Shahid Sadoughi University of Medical Sciences, Yazd, IranDept. of Medical Chemistry, Medical University of Vienna, Vienna, AustriaDept. of Animal Science, University of Applied Science and Technology, Yazd, IranDept. of Medical Genetic, National Institute for Genetic Engineering and Biotechnology (NIGEB), Tehran, IranBackground: A significant role of Renin-angiotensin system (RAS) genetic variants in the pathogenesis of essential hypertension and cardiovascular diseases has been proved. This study aimed to develop a new, fast and cheap method for the simultaneous detection of two missense single nucleotide polymorphisms (T207M or rs4762 and M268T orrs699) of angiotensinogen (AGT) in single-step Multiplex Hexa-Primer Amplification Refractory Mutation System - polymerase chain reaction (H-ARMS-PCR). Methods: In this case-control study, 148 patients with coronary artery disease (CAD) and 135 controls were included. The patients were referred to cardiac centers in Afshar Hospital (Yazd, Iran) from 2012 to 2015. Two sets of inner primer (for each SNP) and one set outer primer pairs were designed for genotyping of rs4762 and rs699 in single tube H-ARMS-PCR. Direct sequencing of all samples was also performed to assess the accuracy of this method. DNA sequencing method validated the results of single tube H-ARMS-PCR. Results: We found full accordance for genotype adscription by sequencing method. The frequency of the AGT T521 and C702 alleles was significantly higher in CAD patients than in the control group (OR: 0.551, 95% CI: 0.359-0.846, P=0.008 and OR: 0.629, 95% CI: 0.422-0.936, P=0.028, respectively). Conclusion: This is the first work describing a rapid, low-cost, high-throughput simultaneous detection of rs4762 and rs699 polymorphisms in AGT gene, used in large clinical studies.https://ijph.tums.ac.ir/index.php/ijph/article/view/10085Angiotensinogen geneCoronary artery diseasers4762rs699H-ARMS-PCR |
| spellingShingle | Mehri KHATAMI Mohammad Mehdi HEIDARI Mehdi HADADZADEH Barbara SCHEIBER-MOJDEHKAR Morteza BITARAF SANI Massoud HOUSHMAND Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR Iranian Journal of Public Health Angiotensinogen gene Coronary artery disease rs4762 rs699 H-ARMS-PCR |
| title | Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR |
| title_full | Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR |
| title_fullStr | Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR |
| title_full_unstemmed | Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR |
| title_short | Simultaneous Genotyping of the Rs4762 and Rs699 Polymorphisms in Angiotensinogen Gene and Correlation with Iranian CAD Patients with Novel Hexa-primer ARMS-PCR |
| title_sort | simultaneous genotyping of the rs4762 and rs699 polymorphisms in angiotensinogen gene and correlation with iranian cad patients with novel hexa primer arms pcr |
| topic | Angiotensinogen gene Coronary artery disease rs4762 rs699 H-ARMS-PCR |
| url | https://ijph.tums.ac.ir/index.php/ijph/article/view/10085 |
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