Elli Francesca Marta, de Sanctis Luisa, Bergallo Massimiliano, Maffini Maria Antonia, Pirelli Arianna, Galliano Ilaria, Bordogna Paolo, Arosio Maura, Mantovani Giovanna
Department of Clinical Sciences and Community Health, University of Milan, Milan, Italy.
Department of Public Health and Pediatric Sciences, University of Torino, Regina Margherita Children's Hospital-AOU Cittàdella Salute e dellaScienza, Torino, Italy.
Front Genet. 2019 Sep 18;10:862. doi: 10.3389/fgene.2019.00862. eCollection 2019.
McCune-Albright syndrome (MAS) is a rare congenital disorder characterized by the association of endocrine and nonendocrine anomalies caused by somatic activating variants of . The mosaic state of variants makes the clinical presentation extremely heterogeneous depending on involved tissues. Biological samples bearing a low level of mosaicism frequently lead to false-negative results with an underestimation of causative molecular alterations, and the analysis of biopsies is often needed to obtain a molecular diagnosis. To date, no reliable analytical method for the noninvasive testing of blood is available. This study was aimed at validating a novel and highly sensitive technique, the digital PCR (dPCR), to increase the detection rate of alterations in patients with a clinical suspicion of MAS and, in particular, in blood. We screened different tissues (blood, bone, cutis, ovary, and ovarian cyst) collected from 54 MAS patients by different technical approaches. Considering blood, Sanger was unable to detect mutations, the allele-specific PCR and the co-amplification at lower denaturation temperature had a 9.1% and 18.1% detection rate, respectively, whereas the dPCR reached a 37.8% detection rate. In conclusion, the dPCR resulted in a cost-effective, reliable, and rapid method allowing the selective amplification of low-frequency variants and able to improve mutant allele detection, especially in the blood.
McCune - Albright综合征（MAS）是一种罕见的先天性疾病，其特征是由......的体细胞激活变体引起的内分泌和非内分泌异常相关联。变体的镶嵌状态使得临床表现因受累组织而异，极其多样化。携带低水平镶嵌性的生物样本常常导致假阴性结果，从而低估致病分子改变，因此通常需要对活检组织进行分析以获得分子诊断。迄今为止，尚无可靠的血液无创检测分析方法。本研究旨在验证一种新型且高度灵敏的技术——数字PCR（dPCR），以提高临床怀疑患有MAS的患者，尤其是血液中......改变的检测率。我们通过不同技术方法对54例MAS患者采集的不同组织（血液、骨骼、皮肤、卵巢和卵巢囊肿）进行了筛查。就血液而言，Sanger测序无法检测到突变，等位基因特异性PCR和较低变性温度下的共扩增检测率分别为9.1%和18.1%，而dPCR的检测率达到了37.8%。总之，dPCR是一种经济高效、可靠且快速的方法，能够选择性扩增低频变体，并能提高......突变等位基因的检测率，尤其是在血液中。
