Comparison of ARMS and direct sequencing for detection of EGFR mutation and prediction of EGFR-TKI efficacy between surgery and biopsy tumor tissues in NSCLC patients

Abstract

The relationship between epidermal growth factor receptor (EGFR) mutation status and EGFR-tyrosine kinase inhibitors (TKI) efficacy in non-small cell lung cancer (NSCLC) patients has been well established. However, there is no available standard to define the optimal testing method and specimen type required for the detection of EGFR mutations. In this study, we compare results of ADx-amplification refractory mutation system (ARMS) and direct sequencing for the detection of EGFR mutation and prediction of EGFR-TKI efficacy for surgery and biopsy tumor tissues in 158 NSCLC patients. For 71 surgery samples, there were 13 and 17 positive samples detected by direct sequencing and ARMS, respectively. For 87 biopsy samples, direct sequencing and ADx-ARMS found 15 and 32 positive samples, respectively. For surgery samples, sensitivity of direct sequencing and ARMS was 72.2% (13/18) and 94.4% (17/18), respectively. For the biopsy samples, sensitivity of direct sequencing and ARMS was 44.1% (15/34) and 94.1% (32/34), respectively. For the biopsy and surgery samples, the ORRs for EGFR positive and negative patients detected by direct sequencing were 46.1 versus 16.7 and 66.7 versus 1.1%, respectively. For ADx-ARMS, the ORR for EGFR positive patients was significantly higher than for negative patients (55.6 vs. 5.6%). The median progression-free survival time of patients with EGFR wild type detected by direct sequencing (4.2 months) was significantly longer than that of patients with wild type detected by ARMS (1.7 months). ARMS has a higher sensitivity and specificity than direct sequencing for EGFR detection of mutation in both surgical and biopsy samples, and the results from ARMS are more consistent with the efficacy of EGFR-TKIs treatment.

Analysis of progression-free survival of first-line tyrosine kinase inhibitors in patients with non-small cell lung cancer harboring leu858Arg or exon 19 deletions

“…The mutation analysis was performed by direct sequencing with polymerase chian reaction or with SCORPION technology in combination with an Amplified Refractory Mutation System (ARMS, QIAGEN, Hilden, Germany) or competitive allele-specific TaqMan PCR (Cast-PCR, Applied Biosystems, Foster City, CA) with genomic DNA from paraffin-embedded tissue (Supplement Table S1). [51, 52]. …”

A new primer construction technique that effectively increases amplification of rare mutant templates in samples

“…Direct sequencing of PCR products is highly accurate, but has low mutation detection sensitivity, being able to only detect~20% of mutant alleles in a background of normal alleles [24,25]. Although allele-specific (AS) PCR can increase detection sensitivity by using type-specific primers, false positive rates are high due to nonspecific product formation [26][27][28][29]. In such cases, validation of the PCR product using sequencing is unhelpful as the primer alters the original sequences.…”

Advanced lung adenocarcinoma with spinal cord metastasis successfully treated with second EGFR-TKI treatment: a case report and literature review

“…In this case, we performed ARMS to detect EGFR mutation in a plasma sample and obtained negative results. A previous study compared the results of ARMS and direct sequencing for the detection of an EGFR mutation and prediction of EGFR-TKI efficacy for surgery and biopsy tumor tissues in 158 patients with NSCLC, 12 which showed that ARMS has a higher sensitivity and specificity than direct sequencing for the detection of an EGFR mutation in both surgical and biopsy samples. Moreover, the results from ARMS are more consistent with the efficacy of EGFR-TKI treatment.…”