Direct Comparison of 3 PCR Methods in Detecting EGFR Mutations in Patients with Advanced Non–Small-Cell Lung Cancer

Ikeda, Takaya; Nakamura, Yoichi; Yamaguchi, Hiroyuki; Tomonaga, Nanae; Doi, Seiji; Nakatomi, Katsumi; Iida, Takuya; Motoshima, Kohei; Mizoguchi, Kosuke; Nagayasu, Takeshi; Tsukamoto, Kazuhiro; Kohno, Shigeru

doi:10.1016/j.cllc.2012.01.008

Cited by 11 publications

(11 citation statements)

References 29 publications

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“…27 In the present study, the sensitivity of the mutant-enriched PCR method was not inferior to the PNA-LNA PCR and PCR clamp methods. 29 They also analyzed intra-tumor heterogeneity without considering the pathological patterns. We considered that the intra-tumor heterogeneity could be demonstrated by an analysis in accordance with the pathological pattern of mixed-type adenocarcinoma in the present study.…”

Section: Discussionmentioning

confidence: 99%

Analysis of Intratumor Heterogeneity of EGFR Mutations in Mixed Type Lung Adenocarcinoma

Tomonaga

Nakamura

Yamaguchi

et al. 2013

Clinical Lung Cancer

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2 Micro abstractIntra-tumor heterogeneity of epidermal growth factor receptor (EGFR) mutations in patients with mixed-type lung adenocarcinoma was analyzed according to histological patterns. Intra-tumor heterogeneity of EGFR mutations was detected in 9 of 38 tumors, and there was a significant correlation between heterogeneity of EGFR mutations and smoking history (P<0.043). Clinical Practice Points• EGFR mutations are factors predictive of response to EGFR-TKI treatment. However, lung cancer is thought to be the result of the accumulation of genetic alterations over a long course of exposure to a carcinogen, and the existence of intra-tumor heterogeneity of EGFR mutations remains unclear. Thus, the existence of intra-tumor heterogeneity was analyzed in patients with mixed-type lung adenocarcinoma according to histological patterns.• Intra-tumor heterogeneity of EGFR mutations was detected in 9 of 38 tumors, and a significant correlation between heterogeneity of EGFR 3 mutations and smoking history was found.• Response to EGFR-TKIs might be partial and insufficient in tumors having intra-tumor heterogeneity of EGFR mutations. New treatment strategies for patients with lung adenocarcinoma harboring heterogeneous EGFR mutations are needed.

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Section: Discussionmentioning

confidence: 99%

Analysis of Intratumor Heterogeneity of EGFR Mutations in Mixed Type Lung Adenocarcinoma

Tomonaga

Nakamura

Yamaguchi

et al. 2013

Clinical Lung Cancer

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“…0.1 %) [9]. The reliability of PNA–LNA PCR clamp has been also confirmed in clinical settings, with high sensitivity (97 %) and specificity (100 %) demonstrated in variety of cytological specimens (bronchoscopy samples, sputum, pleural and pericardial effusion) in addition to paraffin-embedded tissues [1, 24, 26]. Accordingly, Yamada et al [24], demonstrated that the PNA–LNA PCR clamp method allowed positive diagnosis in 33.6 % of 122 cytological samples from Asian NSCLC patients .…”

Section: Discussionmentioning

confidence: 96%

“…Previously, we demonstrated that the median concentration of DNA isolated from intrabronchial forceps biopsy is 38.3 ng/μl [19]. However, commercially available in vitro diagnostic real-time PCR-based tests (CE-IVD) specifically designed for of the detection of EGFR activating mutations are not validated to analyse samples with less than 150–800 ng of DNA or 10 % of neoplastic cells [2, 5, 25, 26]. Thus, the development of highly sensitive molecular methods appropriate for more technically demanding samples has become a major focus in lung cancer diagnostics.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, the development of highly sensitive molecular methods appropriate for more technically demanding samples has become a major focus in lung cancer diagnostics. Techniques based on allele-specific amplification or on the inhibition of wt gene amplification and the simultaneous enhancement of mutated gene amplification have proven particularly useful owing to the high specificity, relative simplicity and cost effectiveness [26–28]. …”

Section: Discussionmentioning

confidence: 99%

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EGFR activating mutations detected by different PCR techniques in Caucasian NSCLC patients with CNS metastases: short report

Wojas‐Krawczyk

Sternschuss²,

Krawczyk

et al. 2013

Clin Exp Metastasis

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EGFR mutation testing has become an essential determination to decide treatment options for NSCLC. The mutation analysis is often conducted in samples with low percentage of tumour cells from primary tumour biopsies. There is very little evidence that samples from metastatic tissues are suitable for EGFR testing. We had evaluated the frequency of EGFR mutations with three highly sensitive PCR techniques in formalin-fixed, paraffin-embedded samples of 143 NSCLC patients with central nervous system (CNS) metastases. 32 corresponding primary tumours were also examined. We used PCR followed by DNA fragments length analysis (FLA), ASP–PCR and PNA–LNA PCR clamp techniques. We found 9 (6.29 %) EGFR gene mutations in CNS samples: 3 (2.1 %) in exon 19 and 6 (4.2 %) in exon 21. The full concordance between CNS metastases and primary tumour samples was observed. PCR followed by DNA–FLA and PNA–LNA PCR clamp were sensitive enough to detect exon 19 deletions. Two mutations in exon 21 were detected by ASP–PCR only, one L858R substitution was detected only by PNA–LNA PCR clamp. With respect to sensitivity, PCR followed by DNA–FLA achieved a level of detection of at least 10 % of mutated DNA for exon 19 deletion, as for ASP–PCR it was at least 5 % of mutated DNA for L858R substitution. Higher sensitivity of 1 % of mutated DNA was achieved by PNA–LNA PCR clamp technique for both mutations. The use of different methodological techniques authenticates the negative result of molecular tests.

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“…The techniques currently used to identify EGFR mutations in patients with lung cancer include PCR single-strand conformation polymorphism, 22 high-resolution melting analysis, 23 ARMS technology, 24 mutant-enriched PCR, peptide nucleic acid-locked nucleic acid PCR clamping, 25 and direct sequencing. These methods come with distinct advantages and disadvantages.…”

Section: Discussionmentioning

confidence: 99%

EGFR mutations in patients with lung adenocarcinoma in southwest China: are G719S/A and L861Q more likely detected in tumors derived from smokers?

Wang¹,

Mou²,

Yang³

et al. 2013

LCTT

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Background:The clinical characteristics of epidermal growth factor receptor (EGFR) hotspot mutations, such as deletions in exon 19, substitution of L858R in exon 21, and mutations in exon 20, have been widely reported in nonsmall cell lung cancer. However, the clinical features of other low frequency EGFR mutations in these four exons (especially the relationship with smoking history), eg, substitutions of G719S/A/C in exon 18 and L861Q in exon 21, remain unclear. This study investigated the relationship between G719S/A/C and L861Q mutations (in exon 18 and 21) and smoking history. Methods: Specimens from 194 patients with lung adenocarcinoma were analyzed for EGFR mutations in exons 18-21 by high-resolution melting curve analysis and amplification refractory mutation technology to establish the relationship between G719S/A/C and L861Q mutations and smoking history. Results: Ninety-six of 194 tumors (49.5%) were confirmed to be EGFR mutation-positive. Among these mutations, 71 of 104 (68.3%) were from never smokers, six of 17 (35.3%) were from former smokers, and 19 of 73 (26.0%) were from current smokers (P , 0.001). The mutation rate in heavy smokers (5/23, 21.7%) was significantly lower than in light smokers (20/67, 29.9%) and never smokers (71/104, 68.3%, P , 0.001). Seven low frequency EGFR mutations (four substitutions of G719S, and G719 A, respectively, and three of L861Q in exon 21) were identified. Five of these mutations were derived from smokers (one former light smoker, one current heavy smoker, and three current light smokers). Four of these patients had been treated with tyrosine kinase inhibitors and all had a partial response, with median overall survival (14.5 months) and median progression-free survival (6.8 months), being longer than in patients with similarly staged lung adenocarcinoma without EGFR mutation or treatment with tyrosine kinase inhibitors (6.8 and 3.1 months, respectively, according to data from an as yet unpublished study at our institution). Conclusion:This study provides further evidence that smoking status, include years of smoking and number of cigarettes smoked per day, plays an important role in EGFR mutation in patients with lung adenocarcinoma. Five of seven specimens with G719S/A or L861Q mutations coming from smokers indicates that there may be a relationship between G719S/A or L861Q mutation and smoking history. However, regardless of the influence of smoking, the effectiveness of tyrosine kinase inhibitors was satisfactory in four patients harboring G719S/A and L861Q EGFR mutation.

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Direct Comparison of 3 PCR Methods in Detecting EGFR Mutations in Patients with Advanced Non–Small-Cell Lung Cancer

Cited by 11 publications

References 29 publications

Analysis of Intratumor Heterogeneity of EGFR Mutations in Mixed Type Lung Adenocarcinoma

Analysis of Intratumor Heterogeneity of EGFR Mutations in Mixed Type Lung Adenocarcinoma

EGFR activating mutations detected by different PCR techniques in Caucasian NSCLC patients with CNS metastases: short report

EGFR mutations in patients with lung adenocarcinoma in southwest China: are G719S/A and L861Q more likely detected in tumors derived from smokers?

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