title: Evaluation of a Two-Year Routine Application of Molecular Testing of Thyroid Fine-Needle Aspirations Using a Seven-Gene Panel in a Primary Referral Setting in Germany.
abstract: Major differences with respect to the diagnostic performance of a "ruling in" approach in the presurgical diagnosis of indeterminate thyroid fine-needle aspirations (FNAs) have been reported. Therefore, the aim of this prospective multicenter study was to investigate the specific diagnostic impact of mutation testing using a seven-gene panel in a routine primary referral setting analyzing FNAs from endocrinology and nuclear medicine practices in Germany.
  
  
  
  RNA and DNA was extracted from 564 routine air-dried FNA smears obtained from 64 physicians and cytologically graded by one experienced cytopathologist. PAX8/PPARG and RET/PTC rearrangements were detected by quantitative polymerase chain reaction, while BRAF and RAS mutations were detected by pyrosequencing. Molecular data were compared to histology and follow-up >1 year, which were available for 322/348 patients undergoing surgery and 33/74 patients having follow-up. Histology results were obtained from the local routine pathologists who were blinded to the molecular test results.
  
  
  
  BRAF and RET/PTC mutations were associated with carcinoma in 98% and 100% of samples, respectively. RAS and PAX8/PPARG mutations were associated with carcinoma in 31% and 0% of samples, respectively. Thirty-six percent of the carcinomas were identified by molecular testing in the atypia of undetermined significance/follicular lesion of undetermined significance and follicular neoplasm/suspicious for a follicular neoplasm categories, with malignancy rates of 15% and 17%, respectively. Due to a low percentage of RAS mutation-positive carcinomas in combination with a rather high percentage of RAS mutation-positive benign nodules, the positive predictive values of 41% and 36% in the atypia of undetermined significance/follicular lesion of undetermined significance and follicular neoplasm/suspicious for a follicular neoplasm categories offer only limited diagnostic potential.
  
  
  
  In conclusion, the data suggest that the application of the current seven-gene panel in a routine primary referral setting does not improve the presurgical diagnosis of thyroid FNAs. While the diagnostic relevance of RAS mutations in thyroid tumors needs further investigation, more comprehensive mutation panels with more cancer-specific mutations may improve the presurgical diagnosis of thyroid FNAs.
authors.affiliation: 1 Department of Oncology and Arnie Charbonneau Cancer Institute, Cumming School of Medicine, University of Calgary , Calgary, Canada .
authors.forename: Markus
authors.initials: M
authors.lastname: Eszlinger
authors.affiliation: 2 Divisions of Endocrinology and Nephrology, University of Leipzig , Leipzig, Germany .
authors.forename: Katharina
authors.initials: K
authors.lastname: Böhme
authors.affiliation: 2 Divisions of Endocrinology and Nephrology, University of Leipzig , Leipzig, Germany .
authors.forename: Maha
authors.initials: M
authors.lastname: Ullmann
authors.affiliation: 2 Divisions of Endocrinology and Nephrology, University of Leipzig , Leipzig, Germany .
authors.forename: Fabian
authors.initials: F
authors.lastname: Görke
authors.affiliation: 3 Department of Pathology, University Hospital Halle , Halle, Germany .
authors.forename: Udo
authors.initials: U
authors.lastname: Siebolts
authors.affiliation: 4 Amedes MVZ wagnerstibbe für Laboratoriumsmedizin, Hämostaseologie, Humangenetik und Mikrobiologie , Hannover, Germany .
authors.forename: Anna
authors.initials: A
authors.lastname: Neumann
authors.affiliation: 5 Zentrum für Nuklearmedizin und PET/CT , Bremen, Germany .
authors.forename: Christiane
authors.initials: C
authors.lastname: Franzius
authors.affiliation: 6 Radiologie Hoheluft , Hamburg, Germany .
authors.forename: Sabine
authors.initials: S
authors.lastname: Adam
authors.affiliation: 7 Klinik Dr. Hancken , Stade, Germany .
authors.forename: Thomas
authors.initials: T
authors.lastname: Molwitz
authors.affiliation: 8 Praxis für Nuklearmedizin , PET-CT Zentrum, Wiesbaden, Germany .
authors.forename: Christian
authors.initials: C
authors.lastname: Landvogt
authors.affiliation: 9 Praxis für Nuklearmedizin , Oldenburg, Germany .
authors.forename: Bassam
authors.initials: B
authors.lastname: Amro
authors.affiliation: 10 Institut für Radiologie & Nuklearmedizin , Bremerhaven, Germany .
authors.forename: Anja
authors.initials: A
authors.lastname: Hach
authors.affiliation: 11 Endokrinologie & Diabetologie im Zentrum , Stuttgart, Germany .
authors.forename: Berit
authors.initials: B
authors.lastname: Feldmann
authors.affiliation: 12 Praxisgemeinschaft auf dem Meere , Lüneburg, Germany .
authors.forename: Dieter
authors.initials: D
authors.lastname: Graf
authors.affiliation: 13 Radiologie und Nuklearmedizin in der Gottorpstraße , Oldenburg, Germany .
authors.forename: Antje
authors.initials: A
authors.lastname: Wefer
authors.affiliation: 14 Radiologie Oldenburg, Praxis Stehen und Partner GbR , Oldenburg, Germany .
authors.forename: Rainer
authors.initials: R
authors.lastname: Niemann
authors.affiliation: 15 Amedes MVZ , Hamburg, Germany .
authors.forename: Catharina
authors.initials: C
authors.lastname: Bullmann
authors.affiliation: 16 Nuklearmedizinische Praxis Dr.med.Klaushenke , Cloppenburg, Germany .
authors.forename: Günther
authors.initials: G
authors.lastname: Klaushenke
authors.affiliation: 17 Endokrinologische Gemeinschaftspraxis , Zentrum für Hormon- und Stoffwechselstörungen, Frankfurt/Main, Germany .
authors.forename: Reinhard
authors.initials: R
authors.lastname: Santen
authors.affiliation: 18 Röntgenpraxis Georgstraße , Hannover, Germany .
authors.forename: Gregor
authors.initials: G
authors.lastname: Tönshoff
authors.affiliation: 19 Radiologisch-Nuklearmedizinische Gemeinschaftspraxis , Celle, Germany .
authors.forename: Velimir
authors.initials: V
authors.lastname: Ivancevic
authors.affiliation: 19 Radiologisch-Nuklearmedizinische Gemeinschaftspraxis , Celle, Germany .
authors.forename: Andreas
authors.initials: A
authors.lastname: Kögler
authors.affiliation: 20 MVZ Westerwald GmbH , Dernbach, Germany .
authors.forename: Erhard
authors.initials: E
authors.lastname: Bell
authors.affiliation: 21 Schilddrüsen- und Hormon-Praxis , Gera, Germany .
authors.forename: Bernd
authors.initials: B
authors.lastname: Lorenz
authors.affiliation: 22 Diagnostisches Zentrum Göttingen, Gemeinschaftspraxis für Radiologie und Nuklearmedizin , Göttingen, Germany .
authors.forename: Gerald
authors.initials: G
authors.lastname: Kluge
authors.affiliation: 23 Schilddrüse Stuttgart, Fachärzte für Innere Medizin und Nuklearmedizinische Schilddrüsendiagnostik , Stuttgart, Germany .
authors.forename: Christoph
authors.initials: C
authors.lastname: Hartenstein
authors.affiliation: 24 MVZ wagnerstibbe für Gynäkologie, Reproduktionsmedizin, Zytologie, Pathologie und Innere Medizin GmbH , Einbeck, Germany .
authors.forename: Ilka
authors.initials: I
authors.lastname: Ruschenburg
authors.affiliation: 25 Division of Endocrinology, Department of Medicine, and Departments of Oncology, and Biochemistry and Molecular Biology, and Arnie Charbonneau Cancer Institute, Cumming School of Medicine, University of Calgary , Calgary, Canada .
authors.forename: Ralf
authors.initials: R
authors.lastname: Paschke
mesh_terms: D018263:Adenocarcinoma, Follicular / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D044963:Biopsy, Fine-Needle; D002291:Carcinoma, Papillary / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D005260:Female; D005858:Germany; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D000072636:PAX8 Transcription Factor / Q000235:genetics; D047495:PPAR gamma / Q000235:genetics; D000072081:Patched-1 Receptor / Q000235:genetics; D011446:Prospective Studies; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics; D000077273:Thyroid Cancer, Papillary; D013964:Thyroid Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D013965:Thyroidectomy; D055815:Young Adult; D018631:ras Proteins / Q000235:genetics
chemical_list: D000072636:PAX8 Transcription Factor; C083325:PAX8 protein, human; D047495:PPAR gamma; C000606236:PTCH1 protein, human; D000072081:Patched-1 Receptor; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D018631:ras Proteins
keywords: cancer; cytology; fine-needle aspiration; molecular diagnostics; thyroid