title: ALK rearrangement testing by FISH analysis in non-small-cell lung cancer patients: results of the first italian external quality assurance scheme.
abstract: BACKGROUND
  The Italian Association of Medical Oncology and the Italian Society of Anatomic Pathology and Diagnostic Cytopathology organized an external quality assessment (EQA) scheme for anaplastic lymphoma kinase (ALK) rearrangement by florescence in situ hybridization (FISH) analysis in non-small-cell lung cancer (NSCLC).
  
  
  METHODS
  Sections from tissue microarrays, each including 10 NSCLC samples with known ALK status, were first validated in five referral laboratories and then provided to 37 participating centers. The laboratories were requested to perform the FISH test, using their usual protocols, and to complete the analysis within 3 weeks. By using a predefined scoring system, two points were assigned in case of correct genotype and zero points to false-negative or false-positive results. The threshold value to pass the EQA scheme was set at 18 points. Two rounds were planned.
  
  
  RESULTS
  Thirty-four centers submitted the results within the established deadline. Several errors in the evaluation of genotype (n = 18) were reported, with both false-positive (n = 7) and false-negative (n = 11) results. Test failure occurred in seven cases. Two samples were found to be critical by two referral laboratories and seven participating centers. Twenty-six (70%) laboratories passed the first round and six the second round. Overall, 32 (86%) laboratories passed the ALK EQA scheme.
  
  
  CONCLUSIONS
  The results of this first EQA scheme for ALK testing in NSCLC cancer patients indicate that ALK analysis is performed with adequate quality in most Italian laboratories and highlight the importance of EQA in revealing methodological problems that need to be addressed to further increase the reproducibility of molecular tests.
authors.affiliation: *Center of Predictive Molecular Medicine, Center of Excellence on Aging, University-Foundation, Chieti, Italy; †Division of Pathology and Laboratory Medicine, European Institute of Oncology, Milan, Italy; ‡Division of Anatomic Pathology, San Luigi Hospital and University of Turin, Orbassano, Italy; §Pathology Department, Policlinico di Modena, Modena, Italy; ‖Diagnostic and Laboratory Medicine, INT-Fondazione "G.Pascale", Naples, Italy; ¶Medical Oncology Unit, University Hospital, Parma, Italy; #Department of Medical Oncology, Perugia Hospital, Perugia, Italy; **Division of Medical Oncology, S.G. Moscati Hospital, Avellino, Italy; ††Department of Human Pathology and Oncology, University of Florence, Florence, Italy; ‡‡Pathology Department, Casa di Cura San Pio X and I.R.C.C.S. Policlinico San Donato, Milano, Italy; §§Thoracic Oncology Unit, University of Torino, San Luigi Hospital, Torino, Italy; ‖‖Cell Biology and Biotherapy Unit, INT-Fondazione "G.Pascale", Naples, Italy; and ¶¶Medical Oncology Unit, S. Orsola-Malpighi Hospital, Bologna, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Marchetti
authors.forename: Massimo
authors.initials: M
authors.lastname: Barberis
authors.forename: Mauro
authors.initials: M
authors.lastname: Papotti
authors.forename: Giulio
authors.initials: G
authors.lastname: Rossi
authors.forename: Renato
authors.initials: R
authors.lastname: Franco
authors.forename: Sara
authors.initials: S
authors.lastname: Malatesta
authors.forename: Fiamma
authors.initials: F
authors.lastname: Buttitta
authors.forename: Andrea
authors.initials: A
authors.lastname: Ardizzoni
authors.forename: Lucio
authors.initials: L
authors.lastname: Crinò
authors.forename: Cesare
authors.initials: C
authors.lastname: Gridelli
authors.forename: Gian Luigi
authors.initials: GL
authors.lastname: Taddei
authors.forename: Claudio
authors.initials: C
authors.lastname: Clemente
authors.forename: Giorgio
authors.initials: G
authors.lastname: Scagliotti
authors.forename: Nicola
authors.initials: N
authors.lastname: Normanno
authors.forename: Carmine
authors.initials: C
authors.lastname: Pinto
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D015321:Gene Rearrangement; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods / Q000592:standards; D008175:Lung Neoplasms / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D011786:Quality Control; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D015203:Reproducibility of Results; D046888:Tissue Array Analysis / Q000379:methods / Q000592:standards
chemical_list: C000626173:ALK protein, human; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases