Er Tze-Kiong, Chang Ya-Sian, Yeh Kun-Tu, Chang Tien-Jye, Chang Jan-Gowth
Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Clin Lab. 2010;56(5-6):175-86.
To compare high resolution melting (HRM) and primer extension (PE) for mutation screening of KRAS codons 12 and 13.
DNA samples were isolated from 60 colorectal cancer (CRC) patients. HRM and primer extension analyses were used for mutation screening of KRAS codons 12 and 13.
Both methods can detect KRAS mutations in a DNA mixture containing as little as 5% mutant DNA. The concordant rate between the two methods was 100%. HRM analysis is able to distinguish mutant from wild type, however, it is unable to detect the actual base change. PE analysis needs more procedures, is time-consuming and slightly more expensive but it is able to detect the precise mutation.
This study showed that HRM is a reliable screening method to identify mutations in the KRAS gene, and the PE can be used as a diagnostic method to accurately pinpoint the nature of mutations.
比较高分辨率熔解曲线分析（HRM）和引物延伸（PE）用于KRAS基因第12和13密码子突变筛查的效果。
从60例结直肠癌（CRC）患者中分离DNA样本。采用HRM和引物延伸分析对KRAS基因第12和13密码子进行突变筛查。
两种方法均能检测出含有低至5%突变DNA的DNA混合物中的KRAS突变。两种方法的一致率为100%。HRM分析能够区分突变型和野生型，但无法检测出实际的碱基变化。PE分析需要更多步骤，耗时且成本略高，但能够检测出精确的突变。
本研究表明，HRM是一种可靠的KRAS基因突变筛查方法，而PE可作为一种诊断方法准确确定突变的性质。
