title: Increase EGFR Mutations Detection Rate in Lung Adenocarcinoma by Real-Time PCR Screening Followed by Direct Sequencing.
abstract: BACKGROUND
  Recently, a number of small-molecule tyrosine kinase inhibitors (TKIs) have been developed to target the ATP-binding cleft of the epidermal growth factor receptor (EGFR). The presence of EGFR mutations in non-small cell lung cancer (NSCLC) correlates with the responsiveness to TKIs. Therefore, the identification of EGFR mutations before the administration of TKIs of NSCLC has become important. The aim of the present study was to investigate the occurrence of EGFR mutations in the southern Taiwanese population with NSCLC using a combination of real-time polymerase chain reaction (PCR) kit and direct sequencing.
  
  
  METHODS
  In the present study, DNAs were extracted from 249 cases of formalin-fixed, paraffin-embedded NSCLC samples for clinical EGFR mutational analysis by real-time PCR kit and direct sequencing.
  
  
  RESULTS
  The results showed that the frequency of EGFR mutations is 63% in the southern Taiwanese population. Most of the EGFR mutations are located at exons 19 and 21. In addition, we indicated that a combination of real-time PCR kit and direct sequencing increases the rate of mutation by 4%. Direct sequencing revealed 9 EGFR mutations including 6 reported EGFR mutations and 3 novel EGFR mutations.
  
  
  CONCLUSIONS
  In the present study, we have demonstrated that a combination of real-time PCR kit and direct sequencing increases the detection rate of EGFR mutations. Therefore, our proposed EGFR mutation detection strategy could be applied in clinical settings. In addition, our results indicated the prevalence of EGFR mutational status in the southern Taiwanese population.
authors.affiliation: Departments of *Laboratory Medicine ‡Internal Medicine, Division of Hematology and Oncology ∥Center for Lipid Biosciences, Kaohsiung Medical University Hospital †Department of Medical Laboratory Science and Biotechnology §Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Tze-Kiong
authors.initials: TK
authors.lastname: Er
authors.forename: Chin-Wen
authors.initials: CW
authors.lastname: Lin
authors.forename: Ta-Chih
authors.initials: TC
authors.lastname: Liu
authors.forename: Chih-Chieh
authors.initials: CC
authors.lastname: Chen
authors.forename: Li-Hsuan
authors.initials: LH
authors.lastname: Wang
authors.forename: Li-Ling
authors.initials: LL
authors.lastname: Hsieh
authors.forename: Wen-Chan
authors.initials: WC
authors.lastname: Tsai
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000208:ethnology / Q000235:genetics* / Q000473:pathology; D000077192:Adenocarcinoma of Lung; D044466:Asian People; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000208:ethnology / Q000235:genetics* / Q000473:pathology; D066246:ErbB Receptors / Q000737:chemistry / Q000235:genetics*; D005091:Exons; D005260:Female; D005557:Formaldehyde; D015870:Gene Expression; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000208:ethnology / Q000235:genetics* / Q000473:pathology; D008297:Male; D008958:Models, Molecular; D009154:Mutation; D059645:Mutation Rate; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D016610:Tissue Embedding; D016707:Tissue Fixation
chemical_list: D005557:Formaldehyde; C512478:EGFR protein, human; D066246:ErbB Receptors