Application of digital PCR with chip-in-a-tube format to analyze Adenomatous polyposis coli (APC) somatic mosaicism
Application of digital PCR with chip-in-a-tube format to analyze Adenomatous polyposis coli (APC) somatic mosaicism
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DOI：
10.1016/j.cca.2017.10.015
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发表时间：
2017-12-01
影响因子：
5
通讯作者：
Sugimura, Haruhiko
中科院分区：
文献类型：
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作者：
Kahyo, Tomoaki;Iwaizumi, Moriya;Sugimura, Haruhiko
背景资料：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正在迅速推进。Clarity（TM）dPCR平台采用管中芯片的形式以扩大其应用范围，已被用于确定基因拷贝数。然而，在人类样本中的突变检测，在临床实践中最苛刻的任务，尚未报告使用此platform.Methods：的Clarity（TM）dPCR平台被用来检测体细胞腺瘤性息肉病大肠杆菌嵌合体c.834 + 2 T > C，这已被确定使用下一代测序（NGS）技术在散发性家族性腺瘤性息肉病患者。结果：使用dPCR平台计算的患者外周血中的突变率为13.2%。这与使用NGS测定的结果相似（12.7%）。相反，在健康供体中，突变率<0.1%。此外，它被证实，dPCR产物的大小是与其理论value.Conclusion一致：我们的研究结果表明，dPCR平台与chip-in-a-tube格式是适合的嵌合分析，使dPCR产物大小的验证。
Background: Over the past decade, digital PCR (dPCR) technology has significantly improved, and its application in clinical diagnostics is rapidly advancing. The Clarity (TM) 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.Methods: The Clarity (TM) dPCR platform was used to detect somatic Adenomatous polyposis coli mosaicism c.834 + 2 T > 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.Results: 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.Conclusion: 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.