Comparison Study of the Performance of the QIAGEN EGFR RGQ and EGFR Pyro Assays for Mutation Analysis in Non–Small Cell Lung Cancer

Cushman‐Vokoun, Allison M.; Crowley, Ann M.; Rapp, Sharleen A.; Greiner, Timothy C.

doi:10.1309/ajcpmf26abeoychz

Cited by 12 publications

(7 citation statements)

References 20 publications

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“…24 However, the limit of detection of the commercial kit was 5% for all the tumor samples 25 and EGFR RGQ Amplification Refractory Mutation System Scorpions probe-based real-time PCR assay is more clinically and analytically sensitive. 26 Intriguingly, we could not detect 41 FFPE samples using direct sequencing due to the low sensitivity. Although the sensitivity of real-time PCR is superior to direct sequencing, we found that 10 FFPE samples failed to detect EGFR mutations using real-time PCR.…”

Section: Discussionmentioning

confidence: 92%

Increase EGFR Mutations Detection Rate in Lung Adenocarcinoma by Real-Time PCR Screening Followed by Direct Sequencing

Er¹,

Lin²,

Liu

et al. 2015

Applied Immunohistochemistry &Amp; Molecular Morphology

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Section: Discussionmentioning

confidence: 92%

Increase EGFR Mutations Detection Rate in Lung Adenocarcinoma by Real-Time PCR Screening Followed by Direct Sequencing

Er¹,

Lin²,

Liu

et al. 2015

Applied Immunohistochemistry &Amp; Molecular Morphology

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“…Forty-seven percent and 79% of p.T790M-mutated specimens had MAFs less than 10% and 20%, respectively, which is below the limit of detection for Sanger sequencing. Twenty-four percent of p.T790M-mutated specimens had MAFs of 1–5% which is below the limit of detection for pyrosequencing, high resolution melting analysis or real-time PCR assays such as the therascreen EGFR RGQ PCR Kit and cobas EGFR Mutation Test [32–34]. In addition, these assays require separate runs for the detection of p.T790M and other EGFR mutations and therefore may not be suitable for comprehensive mutational profiling in core biopsy or fine needle aspiration specimens where tumor tissue is often limited.…”

Section: Discussionmentioning

confidence: 99%

Heterogeneity of resistance mutations detectable by next-generation sequencing in TKI-treated lung adenocarcinoma

et al. 2016

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EGFR-mutated lung adenocarcinomas routinely develop resistance to tyrosine kinase inhibitors (TKI). To better characterize the relative frequencies of the resistance mechanisms, we analyzed 48 EGFR-mutated TKI-resistant specimens from 41 patients. Next-generation sequencing of post-treatment specimens detected EGFR p.T790M in 31 (79%) of 39 patients, PIK3CA mutations in 10 (26%), EGFR p.S768_V769delinsIL in one, and KRAS p.G12C in one. Five PIK3CA mutations were outside of codons 542, 545, and 1047. Three of four pre-treatment specimens did not carry the PIK3CA mutation found in the post-treatment sample. Small cell carcinoma transformation was identified in four patients; none had p.T790M, including two where p.T790M was identified in the co-existing adenocarcinoma. In p.T790M-mutated specimens, the allele frequency was less than 5% in 24% of cases. p.T790M allele frequency was usually lower than that of the sensitizing mutation indicating that the resistance mutation was present either in a subset of cells or, if the sensitizing mutation was amplified, in a subset of the sensitizing alleles of a dominant clone. Eight patients had multiple resistance mutations, suggesting either multiple separate resistant clones or a single clone harboring multiple resistance mechanisms. PIK3CA mutations appear to be a more significant resistance mechanism than previously recognized.

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“…This was not detected by direct Sanger sequencing, indicating that T790M occurred as a low level somatic mutation. The Qiagen EGFR RGQ is highly sensitive, and for T790M mutation in particular, the kit can detect mutant tumor alleles as low as 7% . This is unlike Sanger sequencing which generally has a detection limit of ∼20–25% mutant alleles.…”

Section: Discussionmentioning

confidence: 99%

“…The Qiagen EGFR RGQ is highly sensitive, and for T790M mutation in particular, the kit can detect mutant tumor alleles as low as 7%. 13 This is unlike Sanger sequencing which generally has a detection limit of 20-25% mutant alleles. The failure of previous studies to identify such nonsynonymous EGFR mutations could be attributed to the use of the less sensitive Sanger sequencing methods.…”

Section: Ca S E Re P O Rtmentioning

confidence: 99%

Carcinoma showing thymus like elements: Report of a case with EGFR T790M mutation

Rajeshwari

Singh

Nambirajan

et al. 2017

Diagnostic Cytopathology

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Carcinoma showing thymus-like differentiation of the thyroid (CASTLE) is a rare tumor involving the thyroid and perithyroidal soft tissues. It shares morphological, immunohistochemical and molecular similarities with thymic carcinomas. Due to its relatively better prognosis, it needs differentiation from other primary and metastatic tumors of this region. A 40-year-old lady presented with a gradually progressive anterior neck swelling for one year. Imaging showed bulky right and left lobes of thyroid along with a solid soft tissue mass in the pretracheal region. Fine needle aspiration smears showed features of poorly differentiated carcinoma. Total thyroidectomy with excision of the mass revealed histopathological features characteristic of CASTLE, with evidence of thyroiditis in adjoining thyroid. Epidermal growth factor receptor (EGFR) assay revealed presence of EGFR T790M somatic mutation in exon 20. The same was not detectable on direct sequencing. We present a rare case of CASTLE, occurring in association with Hashimoto thyroiditis, with emphasis on cytological features and report for the first time the presence of a low level somatic mutation in EGFR (EGFR T790M mutation).

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Comparison Study of the Performance of the QIAGEN EGFR RGQ and EGFR Pyro Assays for Mutation Analysis in Non–Small Cell Lung Cancer

Abstract: Both the RGQ real-time PCR and Pyro assays adequately detect common EGFR mutations; however, the RGQ system is more clinically and analytically sensitive. Performance characteristics should be considered when evaluating these EGFR mutation assays for clinical adoption.

Cited by 12 publications

References 20 publications

Increase EGFR Mutations Detection Rate in Lung Adenocarcinoma by Real-Time PCR Screening Followed by Direct Sequencing

Increase EGFR Mutations Detection Rate in Lung Adenocarcinoma by Real-Time PCR Screening Followed by Direct Sequencing

Heterogeneity of resistance mutations detectable by next-generation sequencing in TKI-treated lung adenocarcinoma

Carcinoma showing thymus like elements: Report of a case with EGFR T790M mutation

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