title: Detection and comparison of EGFR mutations in matched tumor tissues, cell blocks, pleural effusions, and sera from patients with NSCLC with malignant pleural effusion, by PNA clamping and direct sequencing.
abstract: Peptide nucleic acid (PNA)-mediated real-time PCR clamping has higher sensitivity than conventional direct sequencing for detecting mutations. Pleural effusion and serum may provide good samples in which to detect epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. We studied 37 NSCLC patients with malignant pleural effusion. EGFR mutations were assessed by PNA clamping and direct sequencing using tumor tissues, cell blocks, pleural effusion, and serum. Concordance between PNA clamping and direct sequencing results, and the diagnostic performance of pleural effusion were investigated. The κ coefficients for the two methods were 0.68 (p = 0.0007), 0.91 (p < 0.0001), 0.75 (p < 0.0001) and -0.01 (p = 0.8639) for tissues, cell blocks, pleural effusion, and serum, respectively. The diagnostic performance of pleural effusion compared with the combination of tumor tissue and cell blocks showed 89% sensitivity, 100% specificity, positive predictive value of 100%, and negative predictive value of 95% by PNA clamping, and 67% sensitivity, 90% specificity, positive predictive value of 75%, and negative predictive value of 86% by directing sequencing. A patient in whom an EGFR mutation was identified in pleural effusion only by PNA clamping showed a significant response to EGFR tyrosine kinase inhibitor (EGFR-TKI) treatment. In contrast to the limited role of serum samples, pleural effusion had a diagnostic performance for the detection of EGFR mutations in NSCLC that was comparable to that of tumor tissues and cell blocks. The diagnostic performance of PNA clamping was good compared with that of direct sequencing. A more sensitive and accurate detection of EGFR mutations would benefit patients by allowing a better prediction of the response to EGFR-TKI treatment.
authors.affiliation: Department of Internal Medicine, Uijeongbu St. Mary's Hospital, The Catholic University of Korea, Republic of Korea.
authors.forename: Chang Dong
authors.initials: CD
authors.lastname: Yeo
authors.forename: Jin Woo
authors.initials: JW
authors.lastname: Kim
authors.forename: Kwan Hyoung
authors.initials: KH
authors.lastname: Kim
authors.forename: Jick Hwan
authors.initials: JH
authors.lastname: Ha
authors.forename: Chin Kook
authors.initials: CK
authors.lastname: Rhee
authors.forename: Seung Joon
authors.initials: SJ
authors.lastname: Kim
authors.forename: Young Kyoon
authors.initials: YK
authors.lastname: Kim
authors.forename: Chan Kwon
authors.initials: CK
authors.lastname: Park
authors.forename: Sang Haak
authors.initials: SH
authors.lastname: Lee
authors.forename: Mi Sun
authors.initials: MS
authors.lastname: Park
authors.forename: Hyeon Woo
authors.initials: HW
authors.lastname: Yim
mesh_terms: D000328:Adult; D000368:Aged; D002289:Carcinoma, Non-Small-Cell Lung / Q000097:blood / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D020135:Peptide Nucleic Acids / Q000235:genetics*; D016066:Pleural Effusion, Malignant / Q000097:blood / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D020135:Peptide Nucleic Acids; D047428:Protein Kinase Inhibitors; C512478:EGFR protein, human; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases
keywords: Direct sequencing; EGFR mutation; Lung cancer; Malignant pleural effusion; Peptide nucleic acid-mediated real-time PCR clamping