Startseite Assessment of EGFR mutation status using cell-free DNA from bronchoalveolar lavage fluid
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Assessment of EGFR mutation status using cell-free DNA from bronchoalveolar lavage fluid

  • Sojung Park , Jae Young Hur , Kye Young Lee , Jae Cheol Lee , Jin Kyung Rho , Sun Hwa Shin und Chang-Min Choi EMAIL logo
Veröffentlicht/Copyright: 14. Februar 2017
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Clinical Chemistry and Laboratory Medicine (CCLM)
Aus der Zeitschrift Clinical Chemistry and Laboratory Medicine (CCLM) Band 55 Heft 10

Abstract

Background:

Much attention has been focused on epidermal growth factor receptor (EGFR) mutation testing since the introduction of EGFR-tyrosine kinase inhibitors have improved survival in EGFR-positive lung cancer patients. Liquid biopsy using circulating tumor cells or cell-free DNA (cfDNA) has enabled less invasive testing, but requires a highly sensitive method. To date, liquid biopsy using bronchoalveolar lavage (BAL) fluid has rarely been used.

Methods:

From 20 patients with lung adenocarcinoma, we isolated cfDNA from 20 samples of cell-free BAL fluid and 19 cell-free bronchial washing samples. cfDNA was examined for EGFR mutations using peptide nucleic acid (PNA)-mediated PCR clamping method. In cases where the results from the tumor biopsy and BAL-derived cfDNA test were not consistent, PANAMutyper™ R EGFR kit was used along with PNA clamping-assisted fluorescence melting curve analysis.

Results:

We included 17 patients with advanced stage disease and three with non-advanced stage disease. Tumor biopsy detected EGFR mutations in 12 of the patients. One patient had a p.L858R mutation and a de novo p.T790M mutation. The results from PNA-mediated PCR clamping were 75.0% (9/12) concordant with the tumor biopsy results for EGFR mutation status. PANAMutyper with fluorescence melting curve analysis was performed in three cases, which detected EGFR mutations in two more patients (11/12, 91.7%). EGFR mutations were detected in the cfDNA extracted from two bronchial washing samples.

Conclusions:

cfDNA from BAL fluid could be used for molecular testing of EGFR mutations and identification of p.T790M mutations, with an easily applicable method.

Keywords: bronchoalveolar lavage; epidermal growth factor receptor; non-small-cell lung cancer; cell-free DNA; targeted mutation detection; PNA-mediated PCR clamping
Corresponding author: Chang-Min Choi, MD, PhD, Department of Pulmonary and Critical Care Medicine, University of Ulsan College of Medicine, Asan Medical Center, 88 Olympic-ro 43-gil, Songpa-gu, Seoul 05505, Republic of Korea
aSojung Park and Jae Young Hur contributed equally to this work.

  1. Author contributions: C.M.C. has been identified as a corresponding author of the paper, taking responsibility for the integrity of the work as a whole, from study conception to published article. K.Y.L. and J.C.L. contributed to study conception and data acquisition. S.P. and J.Y.H. had full access to all of the data in the study and take responsibility for the integrity and accuracy of the data analysis and drafting. S.H.S. and J.K.R. contributed to data acquisition. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This study was supported by a grant (2015-467) from Asan Institute for Life Sciences, Asan Medical Center, Seoul, South Korea.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2016-4-12
Accepted: 2017-1-5
Published Online: 2017-2-14
Published in Print: 2017-8-28

©2017 Walter de Gruyter GmbH, Berlin/Boston

Artikel in diesem Heft

  1. Frontmatter
  2. Editorials
  3. Reporting LDL-cholesterol levels in the era of intensive lipid management: a clarion call
  4. The challenges of genetic risk scores for the prediction of coronary heart disease
  5. Reviews
  6. Advanced lipoprotein testing for cardiovascular diseases risk assessment: a review of the novel approaches in lipoprotein profiling
  7. A review of the challenge in measuring and standardizing BCR-ABL1
  8. Mini Review
  9. Challenges in the analysis of epigenetic biomarkers in clinical samples
  10. Opinion Paper
  11. Defining a roadmap for harmonizing quality indicators in Laboratory Medicine: a consensus statement on behalf of the IFCC Working Group “Laboratory Error and Patient Safety” and EFLM Task and Finish Group “Performance specifications for the extra-analytical phases”
  12. Genetics and Molecular Diagnostics
  13. Assessment of EGFR mutation status using cell-free DNA from bronchoalveolar lavage fluid
  14. General Clinical Chemistry and Laboratory Medicine
  15. A survey of patients’ views from eight European countries of interpretive support from Specialists in Laboratory Medicine
  16. Verification of examination procedures in clinical laboratory for imprecision, trueness and diagnostic accuracy according to ISO 15189:2012: a pragmatic approach
  17. Expressing analytical performance from multi-sample evaluation in laboratory EQA
  18. A candidate reference method for serum potassium measurement by inductively coupled plasma mass spectrometry
  19. Practical motives are prominent in test-ordering in the Emergency Department
  20. Technical and clinical validation of the Greiner FC-Mix glycaemia tube
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  25. Novel immunoassays for detection of CUZD1 autoantibodies in serum of patients with inflammatory bowel diseases
  26. Hematology and Coagulation
  27. Critical appraisal of discriminant formulas for distinguishing thalassemia from iron deficiency in patients with microcytic anemia
  28. Reference Values and Biological Variations
  29. Reference ranges of thromboelastometry in healthy full-term and pre-term neonates
  30. Cancer Diagnostics
  31. Immunoparesis in IgM gammopathies as a useful biomarker to predict disease progression
  32. Cardiovascular Diseases
  33. Assessment of the clinical utility of adding common single nucleotide polymorphism genetic scores to classical risk factor algorithms in coronary heart disease risk prediction in UK men
  34. Time and age dependent decrease of NT-proBNP after septal myectomy in hypertrophic obstructive cardiomyopathy
  35. Infectious Diseases
  36. Higher serum caspase-cleaved cytokeratin-18 levels during the first week of sepsis diagnosis in non-survivor patients
  37. Letters to the Editor
  38. Data mining for age-related TSH reference intervals in adulthood
  39. Intra-laboratory variation and its effect on gestational diabetes diagnosis
  40. Evaluation of long-term imprecision of automated complete blood cell count on the Sysmex XN-9000 system
  41. Sensitivity of the Sysmex XN9000 WPC-channel for detection of monoclonal B-cell populations
  42. Evaluation of biotin interference on immunoassays: new data for troponin I, digoxin, NT-Pro-BNP, and progesterone
  43. Stability of procalcitonin in cerebrospinal fluid
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  45. Mass spectrometry vs. immunoassay in clinical and forensic toxicology: qui modus in rebus est?
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https://doi.org/10.1515/cclm-2016-0302
Schlagwörter für diesen Artikel
bronchoalveolar lavage; epidermal growth factor receptor; non-small-cell lung cancer; cell-free DNA; targeted mutation detection; PNA-mediated PCR clamping

Artikel in diesem Heft

  1. Frontmatter
  2. Editorials
  3. Reporting LDL-cholesterol levels in the era of intensive lipid management: a clarion call
  4. The challenges of genetic risk scores for the prediction of coronary heart disease
  5. Reviews
  6. Advanced lipoprotein testing for cardiovascular diseases risk assessment: a review of the novel approaches in lipoprotein profiling
  7. A review of the challenge in measuring and standardizing BCR-ABL1
  8. Mini Review
  9. Challenges in the analysis of epigenetic biomarkers in clinical samples
  10. Opinion Paper
  11. Defining a roadmap for harmonizing quality indicators in Laboratory Medicine: a consensus statement on behalf of the IFCC Working Group “Laboratory Error and Patient Safety” and EFLM Task and Finish Group “Performance specifications for the extra-analytical phases”
  12. Genetics and Molecular Diagnostics
  13. Assessment of EGFR mutation status using cell-free DNA from bronchoalveolar lavage fluid
  14. General Clinical Chemistry and Laboratory Medicine
  15. A survey of patients’ views from eight European countries of interpretive support from Specialists in Laboratory Medicine
  16. Verification of examination procedures in clinical laboratory for imprecision, trueness and diagnostic accuracy according to ISO 15189:2012: a pragmatic approach
  17. Expressing analytical performance from multi-sample evaluation in laboratory EQA
  18. A candidate reference method for serum potassium measurement by inductively coupled plasma mass spectrometry
  19. Practical motives are prominent in test-ordering in the Emergency Department
  20. Technical and clinical validation of the Greiner FC-Mix glycaemia tube
  21. Comparison of pneumatic tube system with manual transport for routine chemistry, hematology, coagulation and blood gas tests
  22. Accuracy of cerebrospinal fluid Aβ1-42 measurements: evaluation of pre-analytical factors using a novel Elecsys immunosassay
  23. Evaluation of cannabinoids concentration and stability in standardized preparations of cannabis tea and cannabis oil by ultra-high performance liquid chromatography tandem mass spectrometry
  24. Analytical performance and diagnostic accuracy of six different faecal calprotectin assays in inflammatory bowel disease
  25. Novel immunoassays for detection of CUZD1 autoantibodies in serum of patients with inflammatory bowel diseases
  26. Hematology and Coagulation
  27. Critical appraisal of discriminant formulas for distinguishing thalassemia from iron deficiency in patients with microcytic anemia
  28. Reference Values and Biological Variations
  29. Reference ranges of thromboelastometry in healthy full-term and pre-term neonates
  30. Cancer Diagnostics
  31. Immunoparesis in IgM gammopathies as a useful biomarker to predict disease progression
  32. Cardiovascular Diseases
  33. Assessment of the clinical utility of adding common single nucleotide polymorphism genetic scores to classical risk factor algorithms in coronary heart disease risk prediction in UK men
  34. Time and age dependent decrease of NT-proBNP after septal myectomy in hypertrophic obstructive cardiomyopathy
  35. Infectious Diseases
  36. Higher serum caspase-cleaved cytokeratin-18 levels during the first week of sepsis diagnosis in non-survivor patients
  37. Letters to the Editor
  38. Data mining for age-related TSH reference intervals in adulthood
  39. Intra-laboratory variation and its effect on gestational diabetes diagnosis
  40. Evaluation of long-term imprecision of automated complete blood cell count on the Sysmex XN-9000 system
  41. Sensitivity of the Sysmex XN9000 WPC-channel for detection of monoclonal B-cell populations
  42. Evaluation of biotin interference on immunoassays: new data for troponin I, digoxin, NT-Pro-BNP, and progesterone
  43. Stability of procalcitonin in cerebrospinal fluid
  44. Between-laboratory analysis of IgG antibodies against Aspergillus fumigatus in paired quality control samples
  45. Mass spectrometry vs. immunoassay in clinical and forensic toxicology: qui modus in rebus est?
  46. Great need for changes in higher education in Greece
  47. A note from the Editor in Chief regarding the Letter to the Editor “Great need for changes in higher education in Greece”
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