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Droplet digital PCR-based EGFR mutation detection with an internal quality control index to determine the quality of DNA
- Source :
- Scientific Reports, Scientific Reports, Vol 8, Iss 1, Pp 1-10 (2018)
- Publication Year :
- 2018
- Publisher :
- Nature Publishing Group UK, 2018.
-
Abstract
- In clinical translational research and molecular in vitro diagnostics, a major challenge in the detection of genetic mutations is overcoming artefactual results caused by the low-quality of formalin-fixed paraffin-embedded tissue (FFPET)-derived DNA (FFPET-DNA). Here, we propose the use of an ‘internal quality control (iQC) index’ as a criterion for judging the minimum quality of DNA for PCR-based analyses. In a pre-clinical study comparing the results from droplet digital PCR-based EGFR mutation test (ddEGFR test) and qPCR-based EGFR mutation test (cobas EGFR test), iQC index ≥ 0.5 (iQC copies ≥ 500, using 3.3 ng of FFPET-DNA [1,000 genome equivalents]) was established, indicating that more than half of the input DNA was amplifiable. Using this criterion, we conducted a retrospective comparative clinical study of the ddEGFR and cobas EGFR tests for the detection of EGFR mutations in non-small cell lung cancer (NSCLC) FFPET-DNA samples. Compared with the cobas EGFR test, the ddEGFR test exhibited superior analytical performance and equivalent or higher clinical performance. Furthermore, iQC index is a reliable indicator of the quality of FFPET-DNA and could be used to prevent incorrect diagnoses arising from low-quality samples.
- Subjects :
- 0301 basic medicine
Oncology
medicine.medical_specialty
Lung Neoplasms
lcsh:Medicine
Biology
medicine.disease_cause
Polymerase Chain Reaction
Article
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Internal medicine
Carcinoma, Non-Small-Cell Lung
medicine
Carcinoma
Biomarkers, Tumor
Humans
Digital polymerase chain reaction
lcsh:Science
Mutation
Multidisciplinary
lcsh:R
Clinical performance
DNA
Reference Standards
medicine.disease
Internal quality
ErbB Receptors
030104 developmental biology
chemistry
Molecular Diagnostic Techniques
Egfr mutation
030220 oncology & carcinogenesis
lcsh:Q
Non small cell
Subjects
Details
- Language :
- English
- ISSN :
- 20452322
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....745e08d042b492d7d7c651eace3086cf