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Updated: Jun 21, 2026
Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015
Dual priming oligonucleotide-based multiplex PCR analysis for detection of BRAFV600E mutation in FNAB samples of
Jin Young Kwak1, Eun-Kyung Kim, Jong-Kee Kim
1Department of Radiology, Research Institute of Radiological Science, Yonsei University College of Medicine, Seoul, Korea.
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.
