Diabetes and Cardio-Metabolic Disorders Laboratory, Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan.
Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad, Pakistan.
Mol Biol Rep. 2019 Feb;46(1):443-449. doi: 10.1007/s11033-018-4493-0. Epub 2018 Nov 26.
Angiotensin II (Ang II: a truncated octapeptide of angiotensinogen, AGT) and 11-β-hydroxylase influence regulation of blood pressure. Dysregulation of Ang II and 11-β-hydroxylase can lead to hypertension and elevate aldosterone levels. Polymorphisms in AGT (encodes AGT) and CYP11B1 (encodes 11-β-hydroxylase) shift the paradigm from physiological to pathological. Currently, various high-throughput techniques are used to genotype these polymorphisms. These techniques require expensive infrastructure and reagents. However, in developing countries, where cost is the main limiting factor, it is not feasible to use expensive techniques. So, the aim of current study was to develop efficient low-cost method for genotyping of cardiovascular disease and hypertension associated polymorphisms of AGT (rs4762, rs5051) and CYP11B1 (rs6410). For this, tetra amplification-refractory mutation system-polymerase chain reaction (T-ARMS-PCR) method was developed and optimized for aforementioned AGT and CYP11B1 gene polymorphisms. Efficiency of T-ARMS-PCR was tested by genotyping 776 human samples. These T-ARMS-PCR assays were also validated by Sanger DNA sequencing, where 100% concordance was found, allowing the efficient use of these T-ARMS-PCR assays for polymorphism genotyping in AGT and CYP11B1 in resource limited settings. T-ARMS-PCR is low-cost, efficient and reliable assay for genotyping of AGT and CYP11B1 gene polymorphisms.
血管紧张素 II（Ang II：血管紧张素原的八肽片段，AGT）和 11-β-羟化酶影响血压调节。Ang II 和 11-β-羟化酶的失调可导致高血压和醛固酮水平升高。AGT（编码 AGT）和 CYP11B1（编码 11-β-羟化酶）的多态性将范式从生理转变为病理。目前，各种高通量技术用于对这些多态性进行基因分型。这些技术需要昂贵的基础设施和试剂。然而，在发展中国家，成本是主要的限制因素，使用昂贵的技术是不可行的。因此，当前研究的目的是开发一种高效、低成本的方法，用于基因分型与心血管疾病和高血压相关的 AGT（rs4762、rs5051）和 CYP11B1（rs6410）多态性。为此，开发并优化了四扩增不可阻挡突变系统-聚合酶链反应（T-ARMS-PCR）方法，用于上述 AGT 和 CYP11B1 基因多态性的基因分型。通过对 776 个人类样本进行基因分型来测试 T-ARMS-PCR 的效率。这些 T-ARMS-PCR 检测也通过 Sanger DNA 测序进行了验证，发现 100%一致，允许在资源有限的情况下有效地使用这些 T-ARMS-PCR 检测方法对 AGT 和 CYP11B1 的多态性进行基因分型。T-ARMS-PCR 是一种低成本、高效和可靠的 AGT 和 CYP11B1 基因多态性基因分型方法。
