title: BRAFV600E mutation testing in fine needle aspirates of thyroid nodules: potential value of real-time PCR.
abstract: The BRAF(V600E) mutation is a valuable adjunctive diagnostic tool to ultrasound (US)-guided fine-needle aspiration (US-FNA). The objective of this study was to investigate the potential value of realtime PCR to detect BRAF(V600E) mutation. This study included 447 thyroid nodules in 420 patients who underwent US-FNA and BRAF(V600E) mutation analysis using dual priming oligonucleotide-based multiplex polymerase chain reaction (DPO-PCR) and real-time PCR. We calculated and compared the diagnostic performances of DPO-PCR and real-time PCR to detect BRAF(V600E) mutation in the thyroid nodules. Receiver operating characteristic (ROC) analysis was used to quantify the cut-off value of the Ct values of BRAF(V600E) mutation on real-time PCR. Optimal thresholds were determined (Youden index). We also compared the diagnostic performances between DPO-PCR and real-time PCR after applying the cut-off value on real-time PCR. Sensitivity, accuracy, and NPV were significantly higher in real-time PCR than DPO-PCR. When the optimal cut-off value of 32.4 at Ct values of BRAF(V600E) mutation was adjusted on real-time PCR, sensitivity was 66.2% and specificity was 100%. Sensitivity, accuracy, and NPV of real-time PCR were also significantly higher than DPO-PCR. In contrast, specificity and PPV were not significantly different between DPO-PCR and real-time PCR. Real time PCR can be a promising diagnostic method in detecting BRAF(V600E) mutation using optimal cut-off value.
authors.affiliation: Department of Radiology, Severance Hospital, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Kwak
authors.forename: Kyung Hwa
authors.initials: KH
authors.lastname: Han
authors.forename: Jung Hyun
authors.initials: JH
authors.lastname: Yoon
authors.forename: Eun-Kyung
authors.initials: EK
authors.lastname: Kim
authors.forename: Hee Jung
authors.initials: HJ
authors.lastname: Moon
authors.forename: Yoo-Li
authors.initials: YL
authors.lastname: Kim
authors.forename: Seo-Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Jong Rak
authors.initials: JR
authors.lastname: Choi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D019943:Amino Acid Substitution / Q000235:genetics*; D044963:Biopsy, Fine-Needle; D004252:DNA Mutational Analysis; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics*; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D012372:ROC Curve; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013961:Thyroid Gland / Q000473:pathology*; D016606:Thyroid Nodule / Q000235:genetics* / Q000473:pathology* / Q000601:surgery; D055815:Young Adult
chemical_list: C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf