Institute of Life Sciences and Biomedicine, Far Eastern Federal University, Vladivostok 690922, Russia.
A.V. Zhirmunsky National Scientific Center of Marine Biology, Far Eastern Branch of Russian Academy of Sciences, Federal University, Vladivostok 690041, Russia.
Genes (Basel). 2022 Oct 25;13(11):1940. doi: 10.3390/genes13111940.
Wilson's disease (WD) is an autosomal recessive inherited disorder of copper metabolism resulting from various mutations in the gene. Despite good knowledge and successful treatment options, WD is a severe disease that leads to disability, destructively affecting the quality of life of patients. Currently, none of the available laboratory tests can be considered universal and specific for the diagnosis of WD. Therefore, the introduction of genetic diagnostic methods that allow for the identification of the root cause at any stage over the course of the disease gave hope for an earlier solution of diagnostic issues in patients with WD.
A method for the genetic diagnosis of WD based on ARMS PCR, DreamTaq Green PCR Master Mix and modified primers has been developed. This method is able to detect 14 mutant alleles: p.His1069Gln, p.Glu1064Lys, p.Met769HisfsTer26, p.Gly710Ser, p.Ser744Pro, p.Ala1135GlnfsTer13, p.Arg778Leu, p.Arg1041Trp, p.Arg616Gln, p.Arg778Gly, p.Trp779*, p.Val834Asp, p.Gly943Ser and p.3222_3243+21del43.
The primers for all mutations were highly specific with an absence of wild-type amplification. All the results were validated by direct DNA Sanger sequencing.
This fast and economical method provides coverage for the identified common mutations, thereby making ARMS PCR analysis using DreamTaq Green PCR Master Mix and modified primers feasible and attractive for large-scale routine use.
威尔逊病（WD）是一种常染色体隐性遗传性铜代谢紊乱疾病，由基因中的各种突变引起。尽管对该病有很好的认识和成功的治疗选择，但 WD 是一种严重的疾病，会导致残疾，严重影响患者的生活质量。目前，尚无一种可用的实验室检测方法被认为对 WD 的诊断具有普遍性和特异性。因此，引入遗传诊断方法，可以在疾病过程的任何阶段识别根本原因，这为 WD 患者的早期诊断问题提供了希望。
我们开发了一种基于 ARMS PCR、DreamTaq Green PCR Master Mix 和改良引物的 WD 遗传诊断方法。该方法能够检测 14 种突变等位基因：p.His1069Gln、p.Glu1064Lys、p.Met769HisfsTer26、p.Gly710Ser、p.Ser744Pro、p.Ala1135GlnfsTer13、p.Arg778Leu、p.Arg1041Trp、p.Arg616Gln、p.Arg778Gly、p.Trp779*、p.Val834Asp、p.Gly943Ser 和 p.3222_3243+21del43。
所有突变的引物均具有高度特异性，不存在野生型扩增。所有结果均通过直接 DNA Sanger 测序进行验证。
这种快速且经济的方法为已识别的常见突变提供了覆盖，从而使 DreamTaq Green PCR Master Mix 和改良引物的 ARMS PCR 分析在大规模常规使用中具有可行性和吸引力。
