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PCR-sequencing is a complementary method to amplification refractory mutation system for EGFR gene mutation analysis
Junchang Jiang1, Chunhua Wang2, Xiaoli Yu1
1Department of Pathology, Sir Run Run Shaw Hospital, Medical School, Zhejiang University, Hangzhou, Zhejiang 310016, China.
Experimental and Molecular Pathology
|October 20, 2015
Summary
Amplification Refractory Mutation System (ARMS) is widely used for EGFR mutation testing. PCR-sequencing can confirm ARMS results in cases with later amplifications, improving diagnostic accuracy for non-small cell lung cancer.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Amplification Refractory Mutation System (ARMS) is a prevalent method for EGFR gene mutation analysis in China.
- Distinguishing low mutation abundance from no mutation relies on cutoff Ct or ΔCt values.
Purpose of the Study:
- To evaluate the utility of PCR-sequencing as a complementary method to confirm ARMS results in non-small cell lung cancer (NSCLC) samples.
- To assess the diagnostic accuracy of ARMS, particularly in cases with ambiguous results.
Main Methods:
- 359 NSCLC samples were analyzed using ARMS.
- Seventeen samples with Ct or ΔCt values exceeding cutoff thresholds were retested by PCR-sequencing.
- Treatment responses to Tyrosine Kinase Inhibitors (TKI) were monitored in specific patient groups.
Main Results:
- ARMS identified various EGFR mutations including exon 19 deletions, L858R, G719X, L861Q, and T790M, along with wild-type cases.
- Of 22 samples initially classified as wild-type but showing later amplification curves, 17 were retested by PCR-sequencing.
- PCR-sequencing confirmed mutations in 3/3 exon 19 deletions, 2/2 L858R, and 4/12 T790M cases with later amplifications.
- Two patients with confirmed exon 19 deletion and L858R mutations responded to TKI therapy.
Conclusions:
- PCR-sequencing serves as a valuable complementary technique to validate ARMS findings, especially in cases exhibiting later amplifications.
- This combined approach enhances the accuracy of EGFR mutation detection in NSCLC.
- Accurate mutation status is crucial for guiding TKI treatment decisions in NSCLC patients.
