Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, U.O.C. Medicina Generale, Milano, Italy.
Instituto de Investigación, Hospital 12 de Octubre, Madrid, Spain.
Mol Genet Metab. 2018 Nov;125(3):295-301. doi: 10.1016/j.ymgme.2018.09.002. Epub 2018 Sep 5.
Genetic variants in promoters and alternative-splicing lesions require to be experimentally tested in order to validate them as causatives of a disease. The digital PCR (dPCR) approach, which is an alternative to the classical qPCR, is an innovative and a more sensitive method for the detection and quantification of nucleic acids. In the present study, we identified four HMBS gene mutations affecting the ubiquitous isoform of porphobilinogen deaminase (PBGD) and established a dPCR protocol which would be able to detect the different transcripts of this gene. With the application of this method, we were able to characterize the functional roles of these four genetic variants, demonstrating that all these mutations were causatives of the non-erythroid variant of the acute intermittent porphyria (AIP) disease.
为了验证遗传变异是否为疾病的致病因素，需要对启动子和可变剪接病变中的遗传变异进行实验检测。数字 PCR（dPCR）是一种替代经典 qPCR 的创新方法，是一种用于检测和定量核酸的更灵敏的方法。在本研究中，我们鉴定了影响普遍存在的卟胆原脱氨酶（PBGD）同种型的四个 HMBS 基因突变，并建立了能够检测该基因不同转录本的 dPCR 方案。通过应用该方法，我们能够描述这四个遗传变异的功能作用，证明这些突变均为非红细胞急性间歇性卟啉症（AIP）疾病的致病因素。
