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Gangmin Han 1 Article
KRAS Mutation Detection in Non-small Cell Lung Cancer Using a Peptide Nucleic Acid-Mediated Polymerase Chain Reaction Clamping Method and Comparative Validation with Next-Generation Sequencing
Boram Lee, Boin Lee, Gangmin Han, Mi Jung Kwon, Joungho Han, Yoon-La Choi
Korean J Pathol. 2014;48(2):100-107.   Published online April 28, 2014
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AbstractAbstract PDF

KRAS is one of commonly mutated genetic "drivers" in non-small cell lung cancers (NSCLCs). Recent studies indicate that patients with KRAS-mutated tumors do not benefit from adjuvant chemotherapy, so there is now a focus on targeting KRAS-mutated NSCLCs. A feasible mutation detection method is required in order to accurately test for KRAS status.


We compared direct Sanger sequencing and the peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method in 134 NSCLCs and explored associations with clinicopathological factors. Next-generation sequencing (NGS) was used to validate the results of discordant cases. To increase the resolution of low-level somatic mutant molecules, PNA-mediated PCR clamping was used for mutant enrichment prior to NGS.


Twenty-one (15.7%) cases were found to have the KRAS mutations using direct sequencing, with two additional cases by the PNA-mediated PCR clamping method. The frequencies of KRAS mutant alleles were 2% and 4%, respectively, using conventional NGS, increasing up to 90% and 89%, using mutant-enriched NGS. The KRAS mutation occurs more frequently in the tumors of smokers (p=.012) and in stage IV tumors (p=.032).


Direct sequencing can accurately detect mutations, but, it is not always possible to obtain a tumor sample with sufficient volume. The PNA-mediated PCR clamping can rapidly provide results with sufficient sensitivity.


Citations to this article as recorded by  
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    Ullas Batra, Shrinidhi Nathany, Mansi Sharma, Amrith BP, Joslia T. Jose, Harkirat Singh, Sakshi Mattoo, Anurag Mehta
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  • PEAC: An Ultrasensitive and Cost-Effective MRD Detection System in Non-small Cell Lung Cancer Using Plasma Specimen
    Jianping Xu, Yue Pu, Rui Lin, Shanshan Xiao, Yingxue Fu, Tao Wang
    Frontiers in Medicine.2022;[Epub]     CrossRef
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    Kshitij RB Singh, Parikipandla Sridevi, Ravindra Pratap Singh
    Engineering Reports.2020;[Epub]     CrossRef
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  • Association with PD-L1 Expression and Clinicopathological Features in 1000 Lung Cancers: A Large Single-Institution Study of Surgically Resected Lung Cancers with a High Prevalence of EGFR Mutation
    Lee, Kim, Sung, Lee, Han, Kim, Choi
    International Journal of Molecular Sciences.2019; 20(19): 4794.     CrossRef
  • Peptide Nucleic Acid Clamping and Direct Sequencing in the Detection of Oncogenic Alterations in Lung Cancer: Systematic Review and Meta-Analysis
    Jae-Uk Song, Jonghoo Lee
    Yonsei Medical Journal.2018; 59(2): 211.     CrossRef
  • Comparative analysis of Adam33 mutations in murine lung cancer cell lines by droplet digital PCR, real-time PCR and Insight Onco™ NGS
    Soo-Jin Kim, Eunhee Kim, Kyung-Taek Rim
    Molecular & Cellular Toxicology.2018; 14(2): 221.     CrossRef
  • KRASmutation analysis by next-generation sequencing in endoscopic ultrasound-guided sampling for solid liver masses
    Hyun Jong Choi, Jong Ho Moon, Hee Kyung Kim, Yun Nah Lee, Tae Hoon Lee, Sang-Woo Cha, Young Deok Cho, Sang-Heum Park
    Journal of Gastroenterology and Hepatology.2017; 32(1): 154.     CrossRef
  • Molecular Testing of Lung Cancers
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    Journal of Pathology and Translational Medicine.2017; 51(3): 242.     CrossRef
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    Anjali Gupta, Anuradha Mishra, Nidhi Puri
    Journal of Biotechnology.2017; 259: 148.     CrossRef
  • Application of Peptide Nucleic Acid-based Assays Toward Detection of Somatic Mosaicism
    Christopher S Hong, Chunzhang Yang, Zhengping Zhuang
    Molecular Therapy - Nucleic Acids.2016; 5: e314.     CrossRef
  • Detecting Primary KIT Mutations in Presurgical Plasma of Patients with Gastrointestinal Stromal Tumor
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    Molecular Diagnosis & Therapy.2016; 20(4): 347.     CrossRef
  • Transformation to Small Cell Lung Cancer of Pulmonary Adenocarcinoma: Clinicopathologic Analysis of Six Cases
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  • Clinicopathologic characteristics of EGFR, KRAS, and ALK alterations in 6,595 lung cancers
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    Guhyun Kang, Byung Noe Bae, Byeong Seok Sohn, Jung-Soo Pyo, Gu Hyum Kang, Kyoung-Mee Kim
    Targeted Oncology.2015; 10(4): 597.     CrossRef
  • Low Frequency of KRAS Mutation in Pancreatic Ductal Adenocarcinomas in Korean Patients and Its Prognostic Value
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    Pancreas.2015; 44(3): 484.     CrossRef

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