title: EGFRmutation detection on lung cancer cytological specimens by the novel fully automated PCR-based IdyllaEGFRMutation Assay.
abstract: OBJECTIVE
  In everyday practice, epidermal growth factor receptor (EGFR) testing is centralised in referral laboratories that receive paucicellular cytological specimens. Ideally,EGFRtesting should be carried out in the centre where the patient is diagnosed such that the most cellular slide can be selected from in-house collected cytological material. However, available techniques are little standardised and difficult to be implemented in settings with little expertise in molecular testing. The IdyllaEGFRprototype assay is a rapid and fully automated test which may easily be adopted by a wider number of pathological centres. This study assessed whether an IdyllaEGFRprototype assay can be reliably applied to cytological lung cancer specimens.
  
  
  METHODS
  The limit of detection (LOD) of the IdyllaEGFRprototype assay was assessed by cell line dilution studies. A total of 10 ng was directly placed inside an IdyllaEGFRprototype assay cartridge. Idylla results were compared with fragment length (exon 19 del) and Taqman assays.
  
  
  RESULTS
  The IdyllaEGFRprototype assay showed an LOD of 1% mutant allele and yielded valid results in 74/76 (97.3%) samples, detecting all the mutant cases (n=32) identified by standard techniques; in addition, Idylla detected two low abundanceEGFRexon 19 deletions and two G719X exon 18 point mutations, not covered by our standard reference method.
  
  
  CONCLUSIONS
  IdyllaEGFRprototype assay is sensitive on extracted DNA and can reliably be applied on cytological samples, enabling implementation ofEGFRtesting even in less experienced diagnostic units.
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: De Luca
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Gragnano
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Pasquale
authors.initials: P
authors.lastname: Pisapia
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Vigliar
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Umberto
authors.initials: U
authors.lastname: Malapelle
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Claudio
authors.identifier: http://orcid.org/0000-0002-7479-6457
authors.initials: C
authors.lastname: Bellevicine
authors.affiliation: Department of Public Health, University of Naples Federico II, Naples, Italy.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Troncone
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics*; D018773:Genes, erbB-1; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: CYTOLOGY; EGFR; LUNG CANCER; MOLECULAR PATHOLOGY