K-ras mutation detection using a novel amplification refractory mutation system-based quantitative polymerase chain reaction with locked nucleic acid blocker in plasma among colorectal cancer patients

突变 结直肠癌 核酸 聚合酶链反应 数字聚合酶链反应 锁核酸 突变率 耐火材料(行星科学) 分子生物学 癌症研究 癌症 生物 DNA 遗传学 基因 寡核苷酸 天体生物学
作者
Lishan Min,Zhaozheng Zheng,Fuchu Qian,Guiyang Zhang,Yingrong Chen,Jing Zhong,Zhihong Ma,Licheng Dai,Weiqin Jiang
出处
期刊:Chinese journal of experimental surgery 卷期号:34 (11): 1974-1977
标识
DOI:10.3760/cma.j.issn.1001-9030.2017.11.051
摘要

Objective To determine whether plasma circullating cell-free DNA (cfDNA) is a viable alternative approach for K-ras mutation testing in colorectal cancer (CRC) patients. Methods The study included 155 CRC patients. An a novel amplification refractory mutation system-based quantitative polymerase chain reaction with locked nucleic acid (LNA-ARMS-PCR) was developed to assess the K-ras mutation in preoperative plasma and paired tumor tissues. Firstly intra-tumoral heterogeneity analysis was used to determine whether it was a robust mutation detection method for the detection of low-abundance K-ras mutations. Results The newly built LNA-ARMS-PCR was a robust mutation detection method to escape the most false negative caused by intra-tumoral heterogeneity. The positive consensus rate and negative consensus rate was 35.3% and 100.0%, between plasma and tumor tissue K-ras mutation detection, respectively. The majority of K-ras mutations detected in tumors were also found in plasma(15/18 (83.3%)) in patients of stage Ⅳ. The accordance of plasma and tumor mutation are strongly correlated to TNM stage and cfDNA level. Conclusion K-ras analysis in plasma cfDNA is only a viable alternative to tissue analysis in mCRC (stage Ⅳ). Quantitative levels of cfDNA and plasma mutation positivity are strongly correlated. Key words: Colorectal cancer; K-ras; Locked nucleic acid; Amplification refractory mutation system
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