Research Papers:
Rapid and accurate detection of KRAS mutations in colorectal cancers using the isothermal-based optical sensor for companion diagnostics
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Abstract
Choong Eun Jin1,*, Seung-Seop Yeom2,*, Bonhan Koo1, Tae Yoon Lee3, Jeong Hoon Lee4, Yong Shin1 and Seok-Byung Lim2
1Department of Convergence Medicine, Asan Medical Center, University of Ulsan College of Medicine, Biomedical Engineering Research Center, Asan Institute of Life Sciences, Asan Medical Center, Seoul, Republic of Korea
2Division of Colon and Rectal Surgery, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea
3Department of Technology Education, Chungnam National University, Daejeon, Republic of Korea
4Department of Electrical Engineering, Kwangwoon University, Seoul, Republic of Korea
*These authors contributed equally to this work
Correspondence to:
Yong Shin, email: shinyongno1@gmail.com
Seok-Byung Lim, email: sblim@amc.seoul.kr
Keywords: KRAS mutations, colorectal cancer, companion diagnostics, silicon bio-photonic sensor, rapid isothermal DNA amplification
Received: June 08, 2017 Accepted: July 25, 2017 Published: August 08, 2017
ABSTRACT
Although KRAS mutational status testing is becoming a companion diagnostic tool for managing patients with colorectal cancer (CRC), there are still several difficulties when analyzing KRAS mutations using the existing assays, particularly with regard to low sensitivity, its time-consuming, and the need for large instruments. We developed a rapid, sensitive, and specific mutation detection assay based on the bio-photonic sensor termed ISAD (isothermal solid-phase amplification/detection), and used it to analyze KRAS gene mutations in human clinical samples. To validate the ISAD-KRAS assay for use in clinical diagnostics, we examined for hotspot KRAS mutations (codon 12 and codon 13) in 70 CRC specimens using PCR and direct sequencing methods. In a serial dilution study, ISAD-KRAS could detect mutations in a sample containing only 1% of the mutant allele in a mixture of wild-type DNA, whereas both PCR and direct sequencing methods could detect mutations in a sample containing approximately 30% of mutant cells. The results of the ISAD-KRAS assay from 70 clinical samples matched those from PCR and direct sequencing, except in 5 cases, wherein ISAD-KRAS could detect mutations that were not detected by PCR and direct sequencing. We also found that the sensitivity and specificity of ISAD-KRAS were 100% within 30 min. The ISAD-KRAS assay provides a rapid, highly sensitive, and label-free method for KRAS mutation testing, and can serve as a robust and near patient testing approach for the rapid detection of patients most likely to respond to anti-EGFR drugs.
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PII: 20038