title: Direct sequencing in cytological specimens as a useful strategy for detecting EGFR mutations in non-small cell lung cancer patients.
abstract: BACKGROUND
  New therapeutics targeting epidermal growth factor receptor (EGFR) have significantly improved tumor responses to therapy in non-small cell lung cancer (NSCLC) patients. Molecular testing for EGFR mutations informs important therapeutic decisions in clinical practice. In this study, we sought to validate the clinical relevance of sequencing-based EGFR mutation testing combined with cytological analysis using body fluid specimens.
  
  
  METHODS
  Two NSCLC cell lines were used in sensitivity analyses. In addition, we performed cytological analyses and directly sequencing of exons 18-21, for 32 specimens. The absence of EGFR mutations determined by direct sequencing in 14 specimens was confirmed by real-time PCR. Changes made to patients' therapeutic strategies after reports of EGFR mutation status were investigated by querying electronic medical records.
  
  
  RESULTS
  Sensitivity studies showed that detection of in-frame deletions in exon 19 and point mutations in exon 21 was possible in specimens containing 10% and 5% mutant DNA, respectively. In clinical practice, EGFR mutations were detected in 18 of 32 specimens (56.3%). Twelve patients with EGFR mutations detected by direct sequencing were started on treatment with EGFR tyrosine kinase inhibitor (TKI) after reports of EGFR mutation. EGFR-TKI therapy was discontinued for two patients with TKI-resistant T790M mutation. The results of real-time PCR were consistent with those of direct sequencing in 13 of 14 specimens (92.9%) in which no mutation was detected by direct sequencing.
  
  
  CONCLUSIONS
  Combined direct sequencing and cytological analysis of body fluid specimen might be clinically useful and sensitive test for the detection of EGFR mutations in NSCLC patients.
authors: Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Seoul, Reblic of Korea.
authors: Min-Chul
authors: MC
authors: Cho
authors: Chang-Min
authors: CM
authors: Choi
authors: Sollip
authors: S
authors: Kim
authors: Seongsoo
authors: S
authors: Jang
authors: Sejin
authors: S
authors: Jang
authors: Chan-Jeoung
authors: CJ
authors: Park
authors: Hyun-Sook
authors: HS
authors: Chi
authors: Michael J
authors: MJ
authors: Peyton
authors: Woochang
authors: W
authors: Lee
authors: Sail
authors: S
authors: Chun
authors: Sang-We
authors: SW
authors: Kim
authors: Won-Ki
authors: WK
authors: Min
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D001826:Body Fluids / Q000378:metabolism*; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D045744:Cell Line, Tumor; D003584:Cytological Techniques; D004252:DNA Mutational Analysis / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D008297:Male; D008875:Middle Aged; D009362:Neoplasm Metastasis; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D066246:ErbB Receptors