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Abstract
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.
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Fig. 1.
Gel electrophoresis result of polymerase chain reaction-based method. Arrows indicate mutation band.
Fig. 2.
Re-testing result of two false negative cases of polymerase chain reaction-based method in training group for exon 19 mutations. Note faint bands underneath to the wild type DNA bands.
Table 1.
EGFR Mutations in Exons 18∼21 Tested by DNA Sequencing in the Training Set
Table 2.
Comparison of EGFR Mutation Detection Results Using the Sequencing and PCR-Based Methods in the Training Set
Table 3.
Comparison of EGFR Mutation Detection Results Using the Sequencing and PCR-Based Methods in the Test Set
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