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KMID : 0982820120110020077
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Journal of Lung Cancer
2012 Volume.11 No. 2 p.77 ~ p.83
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Screening of Epidermal Growth Factor Receptor Gene Mutation in Non-Small Cell Lung Cancer Using a PCR-Based Enzymatic Digestion Method
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Jung Yoo-Jin
Park Sun-Jung
Lee Sae-Bom
Kim Young-Tae
Park Joo-Yeon
Park In-Kyu
Kang Chang-Hyun
Kim Joo-Hyun
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Abstract
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Purpose: We applied a simplified method using polymerase chain reaction (PCR)-based enzymatic digestion for the detection of epidermal growth factor receptor (EGFR) mutation.
Materials and Methods: We selected 74 samples of adenocarcinoma of the lung with EGFR exons 19 and 21 that had been previously sequenced. We designed PCR primers and chose a DNA restriction enzyme. Seventy four additional lung cancer samples were tested as a test set. For test sets, the PCR-based method was performed first, followed by validation of the result by DNA sequencing.
Results: In the first sample group, we found 15 (20.3%) mutations in exon 19, and 9 (12.2%) mutations in exon 21 using the sequencing method. By using the PCR-based method, we were able to identify all of the mutated samples detected by the sequencing method. The PCR-based method also detected mutations in exon 19 in three additional samples and in exon 21 in one additional sample. In the second sample group, by performing the PCR-based method, we found 10 (13.5%) and 7 (9.5%) mutations in exons 19 and 21, respectively. Additional mutations in exon 19 were identified in 2 samples by the sequencing method. However, the sequencing method failed to identify a mutation in exon 21 in one sample.
Conclusion: The sensitivity of the PCR-based enzymatic digestion method seems to be comparable to that of the traditional sequencing method for detecting EGFR mutations. Our method can be widely used as a screening test to select patients who may benefit from EGFR targeted therapy.
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KEYWORD
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Lung neoplasms, EGFR genes, Mutation, Polymerase chain reaction, Restriction mapping
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