Tomoaki Kahyo1, Moriya Iwaizumi2, Hidetaka Yamada3, Hong Tao3, Kiyotaka Kurachi4, Haruhiko Sugimura5. 1. Department of Tumor Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan. Electronic address: kahyo@hama-med.ac.jp. 2. First Department of Medicine, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan. 3. Department of Tumor Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan. 4. Second Department of Surgery, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan. 5. Department of Tumor Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu, Shizuoka 431-3192, Japan. Electronic address: hsugimur@hama-med.ac.jp.
Abstract
BACKGROUND: 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. METHODS: 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. 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.
BACKGROUND: 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. METHODS: 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. 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.
Authors: Qitao Zhan; Liya Wang; Xiangrong Xu; Yan Sun; Lejun Li; Xuchen Qi; Feng Chen; Xiaoming Wei; Michael L Raff; Ping Yu; Fan Jin Journal: Front Genet Date: 2020-03-04 Impact factor: 4.599