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
Korean Journal of Pathology
;
: 100-107, 2014.
Article
in English
| WPRIM
| ID: wpr-185138
ABSTRACT
BACKGROUND:
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.METHODS:
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.RESULTS:
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).CONCLUSIONS:
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Polymerase Chain Reaction
/
Chemotherapy, Adjuvant
/
Carcinoma, Non-Small-Cell Lung
/
Constriction
/
Peptide Nucleic Acids
/
Alleles
/
Lung Neoplasms
Type of study:
Diagnostic study
Limits:
Humans
Language:
English
Journal:
Korean Journal of Pathology
Year:
2014
Type:
Article
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