Multiplex digital PCR assay to detect multiple KRAS and GNAS mutations associated with pancreatic carcinogenesis from minimal specimen amounts

Digital PCR (dPCR) allows for highly sensitive quantification of low-frequency mutations and facilitates early detection of cancer. However, low throughput targeting of single hotspots in dPCR hinders variant specification when multiple probes are used. Here we developed a dPCR method to simultaneously identify major variants related to pancreatic carcinogenesis. Using a 2-D plot of droplet fluorescence under the optimized concentration of two fluorescent probe pools, we determined the absolute quantification of different KRAS and GNAS variants.

Source: Journal of Molecular Diagnostics - March 23, 2023 Category: Pathology Authors: Maeda Chiho, Yusuke Ono, Akihiro Hayashi, Kenji Takahashi, Kenzui Taniue, Rika Kakisaka, Miyuki Mori, Takahiro Ishii, Hiroki Sato, Tetsuhiro Okada, Hidemasa Kawabata, Takuma Goto, Nobue Tamamura, Yuko Omori, Kuniyuki Takahashi, Akio Katanuma, Hidenori Kar Tags: Regular Article Source Type: research