scholarly journals A CRISPR Test for Rapidly and Sensitively Detecting Circulating EGFR Mutations

Diagnostics ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 114 ◽  
Author(s):  
Jen-Hui Tsou ◽  
Qixin Leng ◽  
Feng Jiang
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Limit Of Detection ◽  
Digital Pcr ◽  
Medical Diagnostics ◽  
Egfr Mutations ◽  
Nucleic Acid Detection ◽  
Great Promise ◽  
Plasma Samples ◽  
Lung Cancer Patients

The detection of EGFR mutations in circulating cell-free DNA can enable personalized therapy for cancer. The current techniques for detecting circulating EGFR mutations are expensive and time-consuming with moderate sensitivity. Emerging CRISPR is revolutionizing medical diagnostics and showing a great promise for nucleic acid detection. This study aims to develop CRISPR-Cas12a as a simple test to sensitively detect circulating EGFR mutations in plasma. Serially diluted samples of DNA containing heterozygous EGFR mutations (L858R and T790M) in wild-type genomic DNA are concurrently tested for the mutations by a CRISPR-Cas12a system and droplet digital PCR (ddPCR). The CRISPR-Cas12a system can detect both L858R and T790M with a limit of detection of 0.005% in less than three hours. ddPCR detects the mutations with a limit of detection of 0.05% for more than five hours. Plasma samples of 28 lung cancer patients and 20 cancer-free individuals are tested for the EGFR mutations by CRISPR-Cas12a system and ddPCR. The CRISPR-Cas12a system could detect L858R in plasma of two lung cancer patients whose tissue biopsies are positive for L858R, and one plasma sample of three lung cancer patients whose tissue biopsies are positive for T790M. ddPCR detects L858R in the same two plasm samples, however, does not detect T790M in any of the plasma samples. This proof of principle study demonstrates that the CRISPR-Cas12a system could rapidly and sensitively detect circulating EGFR mutations, and thus, has potential prognostic or therapeutic implications.

scholarly journals Diagnostic Accuracy of Noninvasive Genotyping of EGFR in Lung Cancer Patients by Deep Sequencing of Plasma Cell-Free DNA

2015 ◽  
Vol 61 (9) ◽  
pp. 1191-1196 ◽  
Author(s):  
Junji Uchida ◽  
Kikuya Kato ◽  
Yoji Kukita ◽  
Toru Kumagai ◽  
Kazumi Nishino ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Deep Sequencing ◽  
Limit Of Detection ◽  
Detection System ◽  
Egfr Mutations ◽  
Plasma Dna ◽  
Lung Cancer Patients ◽  
Therapeutic Decisions ◽  
Exon 19

Abstract BACKGROUND Genotyping of EGFR (epidermal growth factor receptor) mutations is indispensable for making therapeutic decisions regarding whether to use EGFR tyrosine kinase inhibitors (TKIs) for lung cancer. Because some cases might pose challenges for biopsy, noninvasive genotyping of EGFR in circulating tumor DNA (ctDNA) would be beneficial for lung cancer treatment. METHODS We developed a detection system for EGFR mutations in ctDNA by use of deep sequencing of plasma DNA. Mutations were searched in >100 000 reads obtained from each exon region. Parameters corresponding to the limit of detection and limit of quantification were used as the thresholds for mutation detection. We conducted a multi-institute prospective study to evaluate the detection system, enrolling 288 non–small cell lung cancer (NSCLC) patients. RESULTS In evaluating the performance of the detection system, we used the genotyping results from biopsy samples as a comparator: diagnostic sensitivity for exon 19 deletions, 50.9% (95% CI 37.9%–63.9%); diagnostic specificity for exon 19 deletions, 98.0% (88.5%–100%); sensitivity for the L858R mutation, 51.9% (38.7%–64.9%); and specificity for L858R, 94.1% (83.5%–98.6%). The overall sensitivities were as follows: all cases, 54.4% (44.8%–63.7%); stages IA–IIIA, 22.2% (11.5%–38.3%); and stages IIIB–IV, 72.7% (60.9%–82.1%). CONCLUSIONS Deep sequencing of plasma DNA can be used for genotyping of EGFR in lung cancer patients. In particular, the high specificity of the system may enable a direct recommendation for EGFR-TKI on the basis of positive results with plasma DNA. Because sensitivity was low in early-stage NSCLC, the detection system is preferred for stage IIIB–IV NSCLC.

scholarly journals Prevalence of EGFR Mutations and Clinico-Pathological Characteristics of Chilean Lung Cancer Patients

2019 ◽  
Vol 20 (1) ◽  
pp. 1-4 ◽  
Author(s):  
Roger Gejman ◽  
Sergio Gonzalez ◽  
Matias Munoz Medel ◽  
Bruno Nervi ◽  
Cesar Sanchez ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Egfr Mutations ◽  
Lung Cancer Patients ◽  
Pathological Characteristics

scholarly journals BRCA1, LMO4, and CtIP mRNA Expression in Erlotinib-Treated Non–Small-Cell Lung Cancer Patients with EGFR Mutations

2013 ◽  
Vol 8 (3) ◽  
pp. 295-300 ◽  
Author(s):  
Niki Karachaliou ◽  
Carlota Costa ◽  
Ana Gimenez-Capitan ◽  
Miguel Angel Molina-Vila ◽  
Jordi Bertran-Alamillo ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Mrna Expression ◽  
Cancer Patients ◽  
Cell Lung Cancer ◽  
Small Cell ◽  
Egfr Mutations ◽  
Small Cell Lung ◽  
Lung Cancer Patients

scholarly journals Detection of EGFR mutations in plasma DNA from lung cancer patients by mass spectrometry genotyping is predictive of tumor EGFR status and response to EGFR inhibitors

Lung Cancer ◽  
2011 ◽  
Vol 73 (1) ◽  
pp. 96-102 ◽  
Author(s):  
Marie Brevet ◽  
Melissa L. Johnson ◽  
Christopher G. Azzoli ◽  
Marc Ladanyi
Keyword(s):  
Mass Spectrometry ◽  
Lung Cancer ◽  
Cancer Patients ◽  
Egfr Inhibitors ◽  
Egfr Mutations ◽  
Plasma Dna ◽  
Lung Cancer Patients

Abstract 2249: Detection of EGFR mutations in circulating cell-free DNA of non-small cell lung cancer patients by next-generation sequencing

2016 ◽  
Author(s):  
Ken Suzawa ◽  
Shuta Tomida ◽  
Takahiro Matsubara ◽  
Kadoaki Ohashi ◽  
Yuho Maki ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Next Generation Sequencing ◽  
Small Cell Lung Cancer ◽  
Cancer Patients ◽  
Small Cell ◽  
Egfr Mutations ◽  
Small Cell Lung ◽  
Lung Cancer Patients ◽  
Free Dna ◽  
Generation Sequencing
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