Fujita Hiroaki, Fujita Toshitsugu, Ishido Keinosuke, Hakamada Kenichi, Fujii Hodaka
Department of Gastroenterological Surgery, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki, 036-8562, Aomori, Japan.
Department of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki, 036-8562, Aomori, Japan.
Biol Methods Protoc. 2024 Oct 1;9(1):bpae071. doi: 10.1093/biomethods/bpae071. eCollection 2024.
Pancreatic cancer is an aggressive malignancy with a poor prognosis. Single-nucleotide mutations in the gene are detected in the majority of patients with pancreatic ductal adenocarcinoma (PDAC), the most common type of pancreatic cancer. Identifying mutations by liquid biopsy could be effective for detecting and recurrent PDAC; however, sensitive and accurate detection remains challenging. We examined the utility of oligoribonucleotide interference-PCR (ORNi-PCR) followed by real-time PCR or droplet digital PCR (ddPCR) for detecting single-nucleotide mutations by liquid biopsy. A model of cell-free DNA was used to demonstrate that the ORNi-PCR-based methods are more sensitive and accurate for detecting mutant DNA than conventional real-time PCR or ddPCR. ORNi-PCR-based methods could be useful for early detection of and recurrent PDAC by liquid biopsy for cancer diagnosis.
胰腺癌是一种侵袭性恶性肿瘤，预后较差。大多数胰腺导管腺癌（PDAC，最常见的胰腺癌类型）患者中可检测到该基因的单核苷酸突变。通过液体活检鉴定这些突变对于检测早期和复发性PDAC可能有效；然而，灵敏且准确的检测仍然具有挑战性。我们研究了寡核糖核苷酸干扰聚合酶链反应（ORNi-PCR）联合实时聚合酶链反应或数字液滴聚合酶链反应（ddPCR）用于通过液体活检检测单核苷酸突变的效用。使用游离DNA模型证明，基于ORNi-PCR的方法在检测突变DNA方面比传统实时聚合酶链反应或ddPCR更灵敏、准确。基于ORNi-PCR的方法可能有助于通过液体活检对癌症进行诊断，以早期检测和复发性PDAC。
