Rapid and accurate detection of KRAS mutations in colorectal cancers using the isothermal-based optical sensor for companion diagnostics

Choong Eun Jin, Seung Seop Yeom, Bonhan Koo, Tae Yoon Lee, Jeong Hoon Lee, Yong Shin, Seok Byung Lim

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)83860-83871
Number of pages12
JournalOncotarget
Volume8
Issue number48
DOIs
Publication statusPublished - 2017

Bibliographical note

Funding Information:
This work was supported by the grant of the Korea Health Technology R & D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (HI16C-0272-010016), and also supported by a grant (W16-712) from the Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of Korea. The biospecimen and data used in this study was provided by Asan Bio-Resource Center, Korea Biobank Network (2016-13(125)).

Publisher Copyright:
© Jin et al.

All Science Journal Classification (ASJC) codes

  • Oncology

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