title: A comparison of ARMS and direct sequencing for EGFR mutation analysis and tyrosine kinase inhibitors treatment prediction in body fluid samples of non-small-cell lung cancer patients.
abstract: BACKGROUND
  Epidermal growth factor receptor (EGFR) mutation is strongly associated with the therapeutic effect of tyrosine kinase inhibitors (TKIs) in patients with non-small-cell lung cancer (NSCLC). Nevertheless, tumor tissue that needed for mutation analysis is frequently unavailable. Body fluid was considered to be a feasible substitute for the analysis, but arising problems in clinical practice such as relatively lower mutation rate and poor clinical correlation are not yet fully resolved.
  
  
  METHODS
  In this study, 50 patients (32 pleural fluids and 18 plasmas) with TKIs therapy experience and with direct sequencing results were selected from 220 patients for further analysis. The EGFR mutation status was re-evaluated by Amplification Refractory Mutation System (ARMS), and the clinical outcomes of TKIs were analyzed retrospectively.
  
  
  RESULTS
  As compared with direct sequencing, 16 positive and 23 negative patients were confirmed by ARMS, and the other 11 former negative patients (6 pleural fluids and 5 plasmas) were redefined as positive, with a fairly well clinical outcome (7 PR, 3 SD, and 1 PD). The objective response rate (ORR) of positive patients was significant, 81.3% (direct sequencing) and 72.7% (ARMS) for pleural fluids, and 80% (ARMS) for plasma. Notably, even reclassified by ARMS, the ORR for negative patients was still relatively high, 60% for pleural fluids and 46.2% for plasma.
  
  
  CONCLUSIONS
  When using body fluids for EGFR mutation analysis, positive result is consistently a good indicator for TKIs therapy, and the predictive effect was no less than that of tumor tissue, no matter what method was employed. However, even reclassified by ARMS, the correlation between negative results and clinical outcome of TKIs was still unsatisfied. The results indicated that false negative mutation still existed, which may be settled by using method with sensitivity to single DNA molecule or by optimizing the extraction procedure with RNA or CTC to ensure adequate amount of tumor-derived nucleic acid for the test.
authors.affiliation: Cancer Center of People's Liberation Army of China, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China. liuyi790114@163.com
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Li
authors.forename: Jian-Jie
authors.initials: JJ
authors.lastname: Li
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Qin
authors.forename: Chuan-Hao
authors.initials: CH
authors.lastname: Tang
authors.forename: Wan-Feng
authors.initials: WF
authors.lastname: Guo
authors.forename: Hai-Xu
authors.initials: HX
authors.lastname: Hu
authors.forename: Sha
authors.initials: S
authors.lastname: Li
authors.forename: Cui-Jing
authors.initials: CJ
authors.lastname: Chen
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.forename: Hong-Jun
authors.initials: HJ
authors.lastname: Gao
authors.forename: Xiao-Qing
authors.initials: XQ
authors.lastname: Liu
mesh_terms: D001483:Base Sequence; D001826:Body Fluids / Q000737:chemistry / Q000378:metabolism; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy* / Q000201:enzymology* / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*
chemical_list: D047428:Protein Kinase Inhibitors; D066246:ErbB Receptors