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Updated: Mar 10, 2026
Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
Published on: October 8, 2015
Immunohistochemistry as an accurate tool for evaluating BRAF-V600E mutation in 130 samples of papillary thyroid
Zakaria Y Abd Elmageed1, Andrew B Sholl2, Koji Tsumagari3
1Department of Surgery, Tulane University School of Medicine, New Orleans, LA; Department of Otolaryngology, Tulane University School of Medicine, New Orleans, LA.
Background:
BRAFV600E mutation has been investigated by immunohistochemistry and has shown high sensitivity and specificity. We aim to investigate the accuracy of immunohistochemistry versus molecular testing of BRAFV600E in papillary thyroid cancer using a large number of polymerase chain reaction-positive BRAFV600E papillary thyroid cancer tissues.
Methods:
We stained 130 formalin-fixed papillary thyroid cancer specimens using the VE1 antibody: 100 BRAFV600E positive and 30 BRAFV600E negative confirmed by PCR. The sensitivity, specificity, and predictive values of the antibody were assessed.
Results:
Immunohistochemistry of BRAFV600E showed 98.0% sensitivity, 93.3% specificity, and positive and negative predictive values of 98.0% and 93.3%, respectively. Of 100 patients with BRAFV600E PCR-positive samples, 97 (97.0%) had cytoplasmic weak (4.0%), moderate (17.0%), and strong (76.0%) immunostaining. In BRAFV600E PCR-negative samples, cytoplasmic staining was not detected in 93.3% (28/30) of papillary thyroid cancer tissues. The receiver operating characteristic curve demonstrated a high validity and comparable immunohistochemistry method (area under the curve = 98.8%) compared with PCR testing.
Conclusion:
The use of VE1 immunohistochemistry for the detection of BRAFV600E in papillary thyroid cancer tissues is a clinically applicable method with high specificity, sensitivity, and positive and negative predictive values. The reliable use of BRAFV600E immunohistochemistry should promulgate the routine use of this method for BRAFV600E detection in papillary thyroid cancer tissues.
