J Lung Cancer.  2012 Dec;11(2):77-83. 10.6058/jlc.2012.11.2.77.

Screening of Epidermal Growth Factor Receptor Gene Mutation in Non-Small Cell Lung Cancer Using a PCR-Based Enzymatic Digestion Method

• Jung YJ
• Park SJ
• Lee SB
• Kim YT
• Park JY
• Park IK
• Kang CH
• Kim JH

• KMID: 2199833
• DOI: http://doi.org/10.6058/jlc.2012.11.2.77

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.
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.

Keyword

Lung neoplasms; EGFR genes; Mutation; Polymerase chain reaction; Restriction mapping

MeSH Terms

Adenocarcinoma
Carcinoma, Non-Small-Cell Lung
Digestion
DNA
DNA Restriction Enzymes
Epidermal Growth Factor
Exons
Genes, erbB-1
Humans
Lung
Lung Neoplasms
Mass Screening
Polymerase Chain Reaction
Receptor, Epidermal Growth Factor
Restriction Mapping
DNA
DNA Restriction Enzymes
Epidermal Growth Factor
Receptor, Epidermal Growth Factor

Figure

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  Fig. 1. Gel electrophoresis result of polymerase chain reaction-based method. Arrows indicate mutation band.

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  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.

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