Ultra–sensitive droplet digital PCR for detecting a low–prevalence somatic GNAQ mutation in Sturge–Weber syndrome

• DOI: 10.1038/srep22985
• Publication Date: 2016-03-09T10:13:17Z
• Authors: Yuri Uchiyama; Mitsuko Nakashima; Satoshi Watanabe; Masakazu Miyajima; Masataka Taguri; Satoko Miyatake; Noriko Miyake; Hirotomo Saitsu; Hiroyuki Mishima; Akira Kinoshita; Hajime Arai; Ko–ichiro Yoshiura; Naomichi Matsumoto
• Keywords: GNAQ; Sturge–Weber syndrome

Abstract Droplet digital PCR (ddPCR), a method for measuring target nucleic acid sequence quantity, is useful determining somatic mutation rates using TaqMan probes. In this study, the detection limit of copy numbers test DNA by ddPCR was determined based on Poisson distribution. Peptide (PNA), which strongly hybridises to lesions, can inhibit amplification PCR. Therefore, combination with PNA and (PNA–ddPCR), could be lowered. We reanalysed GNAQ (c.548G > A) in patients Sturge–Weber syndrome (SWS) PNA–ddPCR. Importantly, among three previously found negative next–generation sequencing, two had mutant allele frequency less than 1%. Furthermore, we were able find same blood leukocyte or saliva derived from four out 40 SWS patients. Vascular anomalies leukocytes originate endothelial cells haemangioblasts, respectively, are both mesodermal origin. may harbour mutation, depending time when acquired. These data suggest possibility diagnosis some SWS.