|
|
|
| Detecting KRAS gene mutations in peripheral blood of colorectal carcinoma with COLD-PCR-HRM |
| LI Yan-li, ZHAO Dong-mei, XU Yu-hong, ZHANG Yan |
| Department of Clinical Lab, Agricultural Reclamation Bureau General Hospital, Harbin 150088, China |
|
|
|
|
Abstract Objective To evaluate the value of COLD-PCR-HRM in detecting KRAS gene mutations in peripheral blood of patients with colorectal carcinoma. Methods DNA from known mutation type was serially diluted into wild-type DNA to the following percentages: 50%, 25%, 10%, 5%, 3%, 2% and 1%. Tasting KRAS gene mutations in different proportions of DNA by conventional PCR-HRM and COLD-PCR-HRM to validate the sensitivity of two testing methods. At the same time, 62 cases of peripheral blood and tumor tissue matching samples were tested by COLD-PCR-HRM for consistency verification. Results The method of PCR-HRM minimum detectable concentration was 3%, and COLD-PCR-HRM was 1%, both methods were significantly higher than that of direct sequencing (P<0.05). KRAS mutations detection in two kinds of sample by COLD-PCR-HRM: 13 cases mutations in serum samples (mutation rate 21.0%, 13/62), 12 cases mutations in tumor tissue (mutation rate 19.4%, 12/62), 9 cases mutations in both two kinds of samples. Mutations in tumor tissue shall prevail, two mutations uniformity was 75.0% (9/12). There were consistency in KRAS mutations in tissue and peripheral blood (κ=0.649; P<0.001). Conclusions COLD-PCR-HRM significantly improved detection sensitivity of KRAS mutations, which provided favorable conditions for its application in low abundance mutation samples.
|
|
|
|
|
|
| [1] |
1MA Cui-cui， 2LI Yang， 3PANG Xiang-peng， 3LOU Ting-ting，1WANG Huan， 1GUO Rui-fang， 4LIU Ya-hang. Changes and correlation analysis of serum leptin and adiponectin expression in patients with gastric and colorectal cancer[J]. Electronic Journal of Metabolism and Nutrition of, 2019, 6(3): 351-356. |
| [2] |
JIN Xin， HU Ren-chong， TIAN Meng-xing， XU Xin-tian，LI Hong-xia， DAI Zhu， YANG Hui. Meta-analysis of the effects of glutamine on postoperative complications， immune function and nutritional status of colorectal cancer[J]. Electronic Journal of Metabolism and Nutrition of, 2019, 6(2): 246-255. |
| [3] |
1WANG Qian, 1LIU Jie, 1WANG Ning, 1QI Hong-bing, 2MO Fu-quan, 1ZHOU Wen-su, 3ZHANG Pei-jun . The clinical significance of survivin mRNA expression in peripheral blood of patients with colorectal cancer[J]. Electronic Journal of Metabolism and Nutrition of, 2019, 6(1): 79-82. |
| [4] |
1TENG Wen-hao, 1WEI Cheng, 2JIANG Wen-wen, 1XIAO Jun, 1ZANG Wei-dong . Application of “single-incision plus one” combined with ERAS for upper rectal and sigmoid colon cancer[J]. Electronic Journal of Metabolism and Nutrition of, 2019, 6(1): 53-57. |
| [5] |
JIANG Wen-juan, XIA Jing, MA Xia. Application of enhanced recovery after sugery in perioperative period of colorectal cancer patients JIANG Wen-juan, XIA Jing, MA Xia[J]. Electronic Journal of Metabolism and Nutrition of, 2018, 5(4): 407-410. |
| [6] |
1LI Dong-yong, 2SUN Yan-bo, 1QI Ying-hua, 1LI Jian-guang, 1WANG Bin, 1LI Zhi-jun, 2CEN Yun-yun, 2XU Peng-yuan. Effect of appetite condition reflex stimulation on tolerance of early enteral nutrition after colorectal-carcinoma surgery[J]. Electronic Journal of Metabolism and Nutrition of, 2018, 5(4): 366-371. |
|
|
|
|
