Dual priming oligonucleotide-based multiplex PCR analysis for detection of BRAFV600E mutation in FNAB samples of thyroid nodules in BRAFV600E mutation-prevalent area

Head Neck. 2010 Apr;32(4):490-8. doi: 10.1002/hed.21210.

Abstract

Background: To evaluate the diagnostic value of dual priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) for the detection of BRAF(V600E) mutations in ultrasound-guided fine-needle aspiration biopsy (US-FNAB) of thyroid nodules.

Methods: Our institutional review board approved this retrospective study, and informed consent was not required from patients. The 130 patients underwent US-FNAB to evaluate BRAF status in thyroid nodules. In FNAB washouts, DPO-based multiplex PCR, direct DNA sequencing, and PCR-restriction fragment length polymorphism (RFLP) were used to detect BRAF(V600E). The diagnostic performance of these methods was calculated. We compared cytologic results by BRAF status.

Results: Diagnostic accuracy and sensitivity were highest when screening with DPO-based multiplex PCR. BRAF(V600E) positivity was a useful marker at thyroid nodules with "suspicious for papillary thyroid carcinoma" or "inadequate" cytological result.

Conclusions: DPO-based multiplex PCR may be an alternative to direct DNA sequencing because of its high sensitivity, high accuracy, and simplicity. BRAF(V600E) may be a useful additional diagnostic marker in BRAF(V600E)-prevalent areas.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Biopsy, Fine-Needle
  • Carcinoma, Papillary / genetics
  • Carcinoma, Papillary / pathology
  • DNA, Neoplasm / genetics
  • Female
  • Humans
  • Immunohistochemistry
  • Male
  • Middle Aged
  • Mutation*
  • Oligonucleotides / genetics
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Proto-Oncogene Proteins B-raf / genetics*
  • Proto-Oncogene Proteins B-raf / metabolism
  • Risk Assessment
  • Sensitivity and Specificity
  • Thyroid Neoplasms / genetics*
  • Thyroid Neoplasms / pathology*
  • Thyroid Nodule / genetics*
  • Thyroid Nodule / pathology*

Substances

  • DNA, Neoplasm
  • Oligonucleotides
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf