PMID- 24193003
OWN - NLM
STAT- MEDLINE
DCOM- 20140626
LR  - 20191210
IS  - 1533-4066 (Electronic)
IS  - 1052-9551 (Linking)
VI  - 22
IP  - 4
DP  - 2013 Dec
TI  - Pyrosequencing for EGFR mutation detection: diagnostic accuracy and clinical 
      implications.
PG  - 196-203
LID - 10.1097/PDM.0b013e3182893f55 [doi]
AB  - EGFR-activating mutations predict responsiveness to EGFR tyrosine kinase 
      inhibitors (TKIs) in non-small cell lung cancer (NSCLC) patients. Mutation 
      screening is crucial to support therapeutic decisions and is commonly conducted 
      using dideoxy sequencing, although its sensitivity is suboptimal in clinical 
      settings. To evaluate the diagnostic performance of pyrosequencing and dideoxy 
      sequencing, we examined EGFR mutation status in a retrospective cohort of 53 
      patients with NSCLCs clinically selected for TKI therapy and whose clinical 
      outcome was available. Moreover, pyrosequencing quantitative results were 
      compared with EGFR amplification data. EGFR mutations were investigated by 
      pyrosequencing and by dideoxy sequencing. Detection rates of both methods were 
      determined by titration assays using NCI-H1975 and HCC-827 cell lines. Increased 
      EGFR copy number was assessed by fluorescence in situ hybridization (FISH). 
      Pyrosequencing showed a higher detection rate than dideoxy sequencing. Tumor 
      control rate of cases with mutant and wild-type EGFR was 86% and 29%, 
      respectively. EGFR amplification was significantly associated with EGFR mutation 
      and a positive correlation between high percentages of mutant alleles and 
      clinical response to TKI was observed. We concluded that pyrosequencing is more 
      sensitive than dideoxy sequencing in mutation screening for EGFR mutations. 
      Detection rate of dideoxy sequencing was suboptimal when low frequencies of 
      mutant alleles or low tumor cell contents were observed. Pyrosequencing enables 
      quantification of mutant alleles that correlates well with increased EGFR copy 
      number assessed by FISH. Pyrosequencing should be used in molecular diagnostic of 
      NSCLC to appropriately select patients who are likely to benefit from TKI 
      therapy.
FAU - Sahnane, Nora
AU  - Sahnane N
AD  - *Department of Surgical and Morphological Sciences, Anatomic Pathology Unit, 
      University of Insubria, Varese, Italy daggerOncology Unit, Ospedale di Circolo, 
      Varese, Italy.
FAU - Gueli, Rossana
AU  - Gueli R
FAU - Tibiletti, Maria G
AU  - Tibiletti MG
FAU - Bernasconi, Barbara
AU  - Bernasconi B
FAU - Stefanoli, Michele
AU  - Stefanoli M
FAU - Franzi, Francesca
AU  - Franzi F
FAU - Pinotti, Graziella
AU  - Pinotti G
FAU - Capella, Carlo
AU  - Capella C
FAU - Furlan, Daniela
AU  - Furlan D
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Diagn Mol Pathol
JT  - Diagnostic molecular pathology : the American journal of surgical pathology, part 
      B
JID - 9204924
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Enzyme Inhibitors)
RN  - EC 2.7.10.1 (EGFR protein, human)
RN  - EC 2.7.10.1 (ErbB Receptors)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Antineoplastic Agents/therapeutic use
MH  - Carcinoma, Non-Small-Cell Lung/drug therapy
MH  - Enzyme Inhibitors/therapeutic use
MH  - ErbB Receptors/*genetics
MH  - Female
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - *Mutation
MH  - Pathology, Molecular/*methods
MH  - Retrospective Studies
MH  - Sequence Analysis, DNA/*methods
MH  - Treatment Outcome
EDAT- 2013/11/07 06:00
MHDA- 2014/06/27 06:00
CRDT- 2013/11/07 06:00
PHST- 2013/11/07 06:00 [entrez]
PHST- 2013/11/07 06:00 [pubmed]
PHST- 2014/06/27 06:00 [medline]
AID - 10.1097/PDM.0b013e3182893f55 [doi]
PST - ppublish
SO  - Diagn Mol Pathol. 2013 Dec;22(4):196-203. doi: 10.1097/PDM.0b013e3182893f55.