Yang Xue-Min, Liang Jian-Ping, Huang Xiao-Juan, Wang Xiang-Rong, Sun Yang, Dong Chen, Cui Ya-Li, Hui Wen-Li
The College of Life Science, Northwest University, Xi'an, 710069, Shanxi, China.
Data Center of Shaanxi Provincial People's Hospital, Xi'an, 710069, Shanxi, China.
Biol Proced Online. 2021 Jan 7;23(1):2. doi: 10.1186/s12575-020-00140-6.
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.
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各种病理生理状况。
在本研究中，通过整合无需DNA提取的直接PCR，设计了一种用于ACE I/D多态性的新型视觉检测方法，该方法使用基于金磁纳米颗粒（GMNPs）的侧向流动分析（LFA）生物传感器。整个检测过程能够在约80分钟内对临床样本进行基因分型。检测限为0.75 ng，使用LFA装置可在5分钟内获得结果。使用直接PCR-LFA系统分析了300份外周血样本，然后通过测序进行验证以确定准确性和可重复性。随后进行了一项临床初步研究，共分析了633份临床样本。
按年龄分组后，我们发现冠心病组患者的基因型与年龄之间存在显著差异。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。
