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논문 기본 정보

자료유형
학술저널
저자정보
Jinyoung Hong (University of Ulsan College of Medicine) Ji Hyun Kim (University of Ulsan College of Medicine) Seungman Park (Seegene Medical Foundation) Sang Gon Lee (Greencross Laboratories) Woochang Lee (University of Ulsan College of Medicine) Sail Chun (University of Ulsan College of Medicine) Won-Ki Min (University of Ulsan College of Medicine)
저널정보
대한임상검사정도관리협회 Journal of Laboratory Medicine And Quality Assurance Journal of Laboratory Medicine And Quality Assurance Vol.43 No.1
발행연도
2021.1
수록면
31 - 36 (6page)

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Background: DNA extracted from mutant cell lines is frequently used as an external quality assessment (EQA) material for genetic testing of solid tumors because it is easy to obtain. However, the proportion of mutations in cell lines is different from that in actual tumor samples. In this study, mixtures of mutant DNA and wild-type DNA mimicking patient samples were analyzed to optimize the amount of mutant DNA in EQA specimens. Methods: Four cell lines harboring the selected mutation were cultured, and genomic DNA was extracted from cultured cells. Wild-type cell line DNA was prepared in the same manner. Diluted samples were prepared by mixing each mutant cell line DNA and wild-type cell line DNA at different ratios. Sanger sequencing of target variants was performed. For reliability, sequencing was repeated three times and read by two readers. The cutoff was based on the lowest proportion of mutant DNA that was determined to be positive in all three experiments. Results: The cutoffs of mutant cell line DNA ratios were 10%, 5%, 25%, and 25% for KRAS G12C, EGFR exon 19 deletion, EGFR T790M, and BRAF V600E, respectively. For the cell lines harboring EGFR T790M and BRAF V600E, the mutant fraction was not 100%. Conclusions: When manufacturing EQA material for solid tumor genetic testing, consistent results can be obtained if the mutant proportion is 10% or more. In addition, the mutant allele frequency of the cell line should be checked in advance to guarantee that EQA materials contain enough mutant DNA.
#Genetic testing #Laboratory proficiency testing #Somatic variant

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참고문헌 (7)

참고문헌 신청
Li MM / 2017 / Standards and guidelines for the interpretation and reporting of sequence variants in cancer: a joint consensus recommendation of the Association for Molecular Pathology, American Society of Clinical Oncology, and College of American Pathologists / J Mol Diagn 19:4~23 google schola The Food and Drug Administration / Companion diagnostics google schola Mitsudomi T / 2005 / Mutations of the epidermal growth factor receptor gene predict prolonged survival after gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence / J Clin Oncol 23:2513~2520 google schola Clinical and Laboratory Standards Institute / 2013 / Design of molecular proficiency testing/external quality assessment;approved guideline / Clinical and Laboratory Standards Institute google schola Tsiatis AC / 2010 / Comparison of Sanger sequencing, pyrosequencing, and melting curve analysis for the detection of KRAS mutations: diagnostic and clinical implications / J Mol Diagn 12:425~432 google schola

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