Department of Tumor Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan.
First Department of Medicine, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan.
Clin Chim Acta. 2017 Dec;475:91-96. doi: 10.1016/j.cca.2017.10.015. Epub 2017 Oct 18.
Over the past decade, digital PCR (dPCR) technology has significantly improved, and its application in clinical diagnostics is rapidly advancing. The Clarity™ dPCR platform, which employs the chip-in-a-tube format to broaden its range of applications, has been used to determine gene copy number. However, detection of mutations in human samples, the most demanding task in clinical practice, has not yet been reported using this platform.
The Clarity™ dPCR platform was used to detect somatic Adenomatous polyposis coli mosaicism c.834+2T>C, which had been identified using next-generation sequencing (NGS) technology in a patient with sporadic familial adenomatous polyposis. In addition, we were able to determine the size of the dPCR product.
The mutation rate in the peripheral blood of the patient calculated using the dPCR platform was 13.2%. This was similar to that determined using NGS (12.7%). In contrast, in healthy donors, the mutation rate was <0.1%. Furthermore, it was confirmed that the dPCR product size was consistent with its theoretical value.
Our results show that the dPCR platform with the chip-in-a-tube format is suitable for the analysis of mosaicism and enables the validation of the dPCR product size.
在过去的十年中，数字 PCR（dPCR）技术得到了显著的改进，其在临床诊断中的应用也在迅速发展。Clarity™ dPCR 平台采用管中芯片的形式来拓宽其应用范围，已被用于检测基因拷贝数。然而，对于临床实践中最具挑战性的人类样本突变检测，尚未有报道使用该平台进行检测。
使用 Clarity™ dPCR 平台检测了散发性家族性腺瘤性息肉病患者中的体细胞 APC 基因 c.834+2T>C 杂合性，该突变是使用下一代测序（NGS）技术在患者中发现的。此外，我们还能够确定 dPCR 产物的大小。
使用 dPCR 平台计算出的患者外周血中的突变率为 13.2%。这与使用 NGS 确定的突变率（12.7%）相似。相比之下，在健康供体中，突变率<0.1%。此外，还证实了 dPCR 产物的大小与其理论值一致。
我们的结果表明，采用管中芯片形式的 dPCR 平台适用于嵌合体分析，并能够验证 dPCR 产物的大小。
