Invader® Assay for Single-Nucleotide Polymorphism Genotyping and Gene Copy Number Evaluation

Mast, Andrea; Arruda, Monika de

doi:10.1385/1-59745-069-3:173

Cited by 14 publications

(14 citation statements)

References 10 publications

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“…Mutational analysis of the EGFR gene was performed using Scorpion technology in combination with the Amplified Refractory Mutation System or polymerase chain reaction-Invader technique, as previously described (22,23).…”

Section: Egfr Mutation Identificationmentioning

confidence: 99%

Impact of metastatic status on the prognosis of EGFR mutation‑positive non‑small cell lung cancer patients treated with first‑generation EGFR‑tyrosine kinase inhibitors

et al. 2017

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Abstract. The aim of the present study was to analyze the impact of metastatic status on the prognosis of epithelial growth factor receptor (EGFR) mutation-positive patients with non-small cell lung cancer (NSCLC) treated with first-generation EGFR-tyrosine kinase inhibitors (TKIs). A total of 178 EGFR mutation-positive patients with stage IIIB-IV and relapsed NSCLC who were treated with gefitinib or erlotinib as the first-line treatment were enrolled in the present study. Metastatic status, progression-free survival (PFS), overall survival (OS) and treatment-response rates were investigated. The association between the number of metastatic organ sites and patient prognosis was also investigated. The median age at the time of treatment was 72 (range, 39-91) years. A total of 168 patients had adenocarcinoma; 156 were treated with gefitinib. Patients with brain metastases, bone metastases, liver metastases and pleural effusion exhibited a significantly reduced PFS and OS time in the univariate analysis, compared with patients without each of these symptoms. In the multivariate analysis, bone metastasis was associated with a poorer PFS (hazard ratio, 2.11; 95% confidence interval, 1.44-3.09; P<0.001) and brain metastasis was associated with a poorer OS (hazard ratio, 2.41; 95% confidence interval, 1.46-3.95; P<0.001). No association was observed between metastatic status and treatment response rates. Higher numbers of different sites of organ metastases were associated with significantly poorer PFS and OS. Bone, brain metastasis and higher numbers of metastatic organ sites are negative prognostic factors for EGFR mutation-positive NSCLC patients treated with first-generation EGFR-TKIs.

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Section: Egfr Mutation Identificationmentioning

confidence: 99%

Impact of metastatic status on the prognosis of EGFR mutation‑positive non‑small cell lung cancer patients treated with first‑generation EGFR‑tyrosine kinase inhibitors

et al. 2017

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“…Polymerase chain reaction (PCR) invader assay (15), direct sequencing or peptide nucleic acid-locked nucleic acid PCR clamp assay (16) was used for analysis of EGFR mutations. Deletions in or near E746-A750 in exon 19 and L858R in exon 21 were regarded as mutation-positive for the purposes of this trial.…”

Section: Assessment Of Tumor Egfr Mutation Statusmentioning

confidence: 99%

Phase II Trial of Erlotinib for Japanese Patients With Previously Treated Non-small-cell Lung Cancer Harboring EGFR Mutations: Results of Lung Oncology Group in Kyushu (LOGiK0803)

Yamada¹,

Takayama²,

Kawakami³

et al. 2013

Japanese Journal of Clinical Oncology

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Objective: Erlotinib has been reported to be useful for treatment of non-small-cell lung cancer harboring mutation of the epidermal growth factor receptor gene EGFR-mt. However, no prospective trial has yet assessed the utility of erlotinib in Japanese patients. Methods: Patients with EGFR-mt (exon 19/21) non-small-cell lung cancer who had previously received one to two chemotherapy regimens were enrolled in this trial. Erlotinib was initially administered at a dose of 150 mg/day orally until disease progression or unacceptable toxicities occurred. The primary endpoint was the objective response rate. Results: Twenty-six patients were enrolled between February 2009 and January 2011. Objective response was observed in 14 patients (53.8%, 95% confidence interval: 33.4 -73.4%), and the disease control rate reached 80.8% (95% confidence interval: 60.7 -93.5%). After a median follow-up time of 17.3 months (range: 5.8 -29.5 months), the median progression-free survival was 9.3 months (95% confidence interval: 7.6 -11.6 months). The median survival time is yet to be determined. Major toxicities were skin disorder and liver dysfunction; most episodes were grade 2 or less, and all were tolerable. Only one patient with grade 3 skin rash discontinued the study. No patients developed interstitial lung disease, and there were no treatment-related deaths. Conclusions: This prospective study is the first to have investigated the usefulness of erlotinib in Japanese patients with previously treated EGFR-mt non-small-cell lung cancer. Although this trial could not meet the primary endpoint, erlotinib was well tolerated and showed clinical benefit such as promising disease control rate or progression-free survival in this population, similar to gefitinib.

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“…The remaining PCR primers were designed by Primer Express 1.5 (Applied Biosystems, Foster City, CA), and the other invader probes and allele probes were designed and synthesized under the reported criteria [Mast and de Arruda, 2006]. The sequences of all primers and probes are listed in Supplementary Table S1 (available online at http://www.interscience.wiley.com/jpages/1059-7794/ suppmat).…”

Section: Pcr-retinamentioning

confidence: 99%

Multiplex PCR-based real-time invader assay (mPCR-RETINA): a novel SNP-based method for detecting allelic asymmetries within copy number variation regions

et al. 2008

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We report the development of a real-time Invader assay combined with multiplex PCR (mPCR-RETINA), an SNP-based approach that can measure the allelic ratio in copy number variation (CNV) regions of a genome. RETINA monitors the real-time fluorescence intensity of each allele during the Invader assay and detects allelic asymmetries caused by genomic duplication/multiplication in heterozygous individuals. By combining mPCR-RETINA and real-time quantitative PCR that detects total copy number, we can estimate the copy number of each allele in CNV regions, which should be useful for investigating the functional significance of allele copy number with disease susceptibilities and drug responses. Also, mPCR-RETINA can efficiently refine the detailed structures of CNV regions. Due to the combination of RETINA with multiplex PCR, mPCR-RETINA requires a very small amount of genomic DNA for analysis (0.1-0.38 ng/locus). Additionally, mPCR-RETINA has clear advantages in its simple protocol and target-specific reaction, even in nonunique regions. We believe mPCR-RETINA will provide a significant contribution to identifying functional alleles in CNV regions.

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Invader® Assay for Single-Nucleotide Polymorphism Genotyping and Gene Copy Number Evaluation

Cited by 14 publications

References 10 publications

Impact of metastatic status on the prognosis of EGFR mutation‑positive non‑small cell lung cancer patients treated with first‑generation EGFR‑tyrosine kinase inhibitors

Impact of metastatic status on the prognosis of EGFR mutation‑positive non‑small cell lung cancer patients treated with first‑generation EGFR‑tyrosine kinase inhibitors

Phase II Trial of Erlotinib for Japanese Patients With Previously Treated Non-small-cell Lung Cancer Harboring EGFR Mutations: Results of Lung Oncology Group in Kyushu (LOGiK0803)

Multiplex PCR-based real-time invader assay (mPCR-RETINA): a novel SNP-based method for detecting allelic asymmetries within copy number variation regions

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