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A Novel PCR Method for Detecting ACE Gene Insertion/Deletion Polymorphisms and its Clinical Application
Biological Procedures Online ( IF 3.7 ) Pub Date : 2021-01-07 , DOI: 10.1186/s12575-020-00140-6
Xue-min Yang , Jian-ping Liang , Xiao-juan Huang , Xiang-rong Wang , Yang Sun , Chen Dong , Ya-li Cui , Wen-li Hui

Angiotensin-converting enzyme (ACE) plays a major role in blood pressure regulation and cardiovascular homeostasis. The wide distribution and multifunctional properties of ACE suggest it’s involvement in various pathophysiological conditions. In this study, a novel visual detection method for ACE I/D polymorphisms was designed by integrating direct PCR without the need for DNA extraction using gold magnetic nanoparticles (GMNPs)-based lateral flow assay (LFA) biosensor. The entire detection procedure could enable the genotyping of clinical samples in about 80 min. The detection limit was 0.75 ng and results could be obtained in 5 min using the LFA device. Three hundred peripheral blood samples were analyzed using the direct PCR-LFA system and then verified by sequencing to determine accuracy and repeatability. A clinical preliminary study was then performed to analyze a total of 633 clinical samples. After grouping based on age, we found a significant difference between the genotypes and the age of patients in the CHD group. The introduction of this method into clinical practice may be helpful for the diagnosis of diseases caused by large fragment gene insertions/deletions.

中文翻译:

检测ACE基因插入/缺失多态性的新型PCR方法及其临床应用

血管紧张素转换酶(ACE)在血压调节和心血管稳态中起主要作用。ACE的广泛分布和多功能特性表明它参与了各种病理生理状况。在这项研究中,通过集成直接PCR而不需要使用基于金磁性纳米颗粒(GMNPs)的侧向流动测定(LFA)生物传感器进行DNA提取,设计了一种ACE I / D多态性的新型视觉检测方法。整个检测过程可以在大约80分钟内完成临床样品的基因分型。检出限为0.75 ng,使用LFA设备可在5分钟内获得结果。使用直接PCR-LFA系统分析了300个外周血样本,然后通过测序进行验证,以确定准确性和可重复性。然后进行了临床初步研究,以分析总共633个临床样品。根据年龄分组后,我们发现冠心病组的基因型和患者年龄之间存在显着差异。将该方法引入临床实践可能有助于诊断由大片段基因插入/缺失引起的疾病。
更新日期:2021-01-08
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