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Assessment of real-time PCR method for detection of EGFR mutation using both supernatant and cell pellet of malignant pleural effusion samples from non-small-cell lung cancer patients

Authors
 Saeam Shin  ;  Juwon Kim  ;  Yoonjung Kim  ;  Sun-Mi Cho  ;  Kyung-A Lee 
Citation
 Clinical Chemistry and Laboratory Medicine, Vol.55(12) : 1962-1969, 2017 
Journal Title
 Clinical Chemistry and Laboratory Medicine 
ISSN
 1434-6621 
Issue Date
2017
MeSH
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung/genetics* ; Carcinoma, Non-Small-Cell Lung/pathology ; DNA Mutational Analysis* ; Female ; Humans ; Lung Neoplasms/genetics* ; Lung Neoplasms/pathology ; Male ; Middle Aged ; Mutation* ; Pleural Effusion, Malignant/genetics* ; Pleural Effusion, Malignant/pathology ; Real-Time Polymerase Chain Reaction* ; Receptor, Epidermal Growth Factor/genetics*
Keywords
EGFR ; cobas ; malignant pleural effusion ; non-small-cell lung cancer ; real-time PCR
Abstract
BACKGROUND: EGFR mutation is an emerging biomarker for treatment selection in non-small-cell lung cancer (NSCLC) patients. However, optimal mutation detection is hindered by complications associated with the biopsy procedure, tumor heterogeneity and limited sensitivity of test methodology. In this study, we evaluated the diagnostic utility of real-time PCR using malignant pleural effusion samples. METHODS: A total of 77 pleural fluid samples from 77 NSCLC patients were tested using the cobas EGFR mutation test (Roche Molecular Systems). Pleural fluid was centrifuged, and separated cell pellets and supernatants were tested in parallel. Results were compared with Sanger sequencing and/or peptide nucleic acid (PNA)-mediated PCR clamping of matched tumor tissue or pleural fluid samples. RESULTS: All samples showed valid real-time PCR results in one or more DNA samples extracted from cell pellets and supernatants. Compared with other molecular methods, the sensitivity of real-time PCR method was 100%. Concordance rate of real-time PCR and Sanger sequencing plus PNA-mediated PCR clamping was 98.7%. CONCLUSIONS: We have confirmed that real-time PCR using pleural fluid had a high concordance rate compared to conventional methods, with no failed samples. Our data demonstrated that the parallel real-time PCR testing using supernatant and cell pellet could offer reliable and robust surrogate strategy when tissue is not available.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/160949
DOI
10.1515/cclm-2016-0851
Appears in Collections:
1. Journal Papers (연구논문) > 1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실)
Yonsei Authors
이경아(Lee, Kyung A)
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https://www.degruyter.com/view/j/cclm.2017.55.issue-12/cclm-2016-0851/cclm-2016-0851.xml
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