title: ALK status testing in non-small-cell lung carcinoma by FISH on ThinPrep slides with cytology material.
abstract: BACKGROUND
  Oncogenic anaplastic lymphoma kinase (ALK) gene rearrangements in non-small-cell lung carcinomas (NSCLC) provide the basis for targeted therapy with crizotinib and other specific ALK inhibitors. Treatment eligibility is conventionally determined by the Food and Drug Administration-approved companion diagnostic fluorescence in situ hybridization (FISH) assay on paraffin-embedded tissue (PET). On limited samples such as fine needle aspiration-derived cytoblocks, FISH for ALK is often uninformative. FISH performed on liquid-based ThinPrep slides (ThinPrep-FISH) may represent a robust alternative.
  
  
  METHODS
  Two hundred thirty cytology samples from 217 patients with advanced NSCLC, including a consecutive series of 179 specimens, were used to generate matched ThinPrep slides and paraffin cytoblocks. The same ThinPrep slides used for cytologic diagnosis were assessed by standard ALK break-apart two-color probe FISH, after etching of tumor areas. Ultrasensitive ALK immunohistochemistry (IHC) on corresponding cytoblocks [D5F3 antibody, OptiView signal amplification] served as the reference data set.
  
  
  RESULTS
  ThinPrep-FISH ALK signals were robust in 228 of 230 cases and not compromised by nuclear truncation inherent in paraffin-embedded tissue-FISH; only two samples displayed no signals. Nine of 178 informative cases (5%) in the consecutive series and 18 of 228 informative cases (7.8%) overall were ALK rearranged by ThinPrep-FISH. In 154 informative matched ThinPrep-FISH and cytoblock-IHC samples, 152 were concordant (10, 6.5% ALK status positive; 142, 92.2% ALK status negative), and two (1.3%) were ThinPrep-FISH positive but IHC negative (sensitivity 100%, specificity 98.6%, overall agreement 98.7%).
  
  
  CONCLUSIONS
  Detection of ALK gene rearrangements in liquid cytology ThinPrep slides derived from patients with NSCLC can be confidently used for clinical ALK molecular testing.
authors.affiliation: *Departments of Molecular Pathology, Cytopathology, and Anatomic Pathology, Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland, Ohio; †Departments of Translational Hematology and Oncology Research, and Solid Tumor Oncology, Taussig Cancer Institute, Cleveland, Ohio; and ‡Respiratory Institute, Cleveland Clinic Foundation, Cleveland, Ohio.
authors.forename: Eugen C
authors.initials: EC
authors.lastname: Minca
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Lanigan
authors.forename: Jordan P
authors.initials: JP
authors.lastname: Reynolds
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Ma
authors.forename: Joseph
authors.initials: J
authors.lastname: Cicenia
authors.forename: Francisco A
authors.initials: FA
authors.lastname: Almeida
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Pennell
authors.forename: Raymond R
authors.initials: RR
authors.lastname: Tubbs
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D000077548:Anaplastic Lymphoma Kinase; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D003581:Cytodiagnosis / Q000379:methods*; D015321:Gene Rearrangement; D006801:Humans; D007124:Immunoenzyme Techniques; D017404:In Situ Hybridization, Fluorescence; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D009367:Neoplasm Staging; D011379:Prognosis; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism*
chemical_list: C000626173:ALK protein, human; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases