DHPLC is a highly sensitive and rapid screening method to detect BRAF(V600E) mutation in papillary thyroid carcinoma.
The BRAF(V600E) mutation has been reported to occur in 30% to 80% of papillary thyroid carcinomas (PTCs). Although direct sequencing is the method most commonly used to identify mutations, this technique is not sensitive enough to accurately detect low level mutation. To determine the optimal diagnostic method for detecting the BRAF(V600E) mutation in PTC, we compared the diagnostic efficacy of four representative detection methods in formalin-fixed paraffin-embedded thyroid tissues obtained from 40 patients diagnosed with PTC. To detect the BRAF(V600E) mutation, we amplified exon 15 of the BRAF gene and performed mutational analysis with direct sequencing, denaturing high-performance liquid chromatography (DHPLC), pyrosequencing and colorimetric assay. The BRAF mutation was detected in 33 cases (82.5%) by DHPLC, 23 cases (57.5%) by direct sequencing, 22 cases (55.0%) by pyrosequencing, and 37 cases (92.5%) by colorimetric assay. The sensitivity, negative predictive value and accuracy of DHPLC were 100%. The specificity and positive predictive values for DHPLC, direct sequencing and pyrosequencing were 100%, and for colorimetric assay they were 14.3% and 83.8%, respectively. The kappa value for DHPLC was a perfect 1.0, which was superior to the other methods. In conclusion, DHPLC is a sensitive, specific and accurate method for detecting the BRAF(V600E) mutation, especially low level mutation, in PTC.
Department of Hospital Pathology, Seoul St. Mary's Hospital, The Catholic University of Korea, Republic of Korea.
Tae Eun
TE
Kim
Eun Sun
ES
Jung
Chan Kwon
CK
Jung
Ja Seong
JS
Bae
Seung Nam
SN
Kim
Gyeong Suk
GS
Kim
Hyoung Nam
HN
Lee
Chang Suk
CS
Kang
Yeong Jin
YJ
Choi
D000328:Adult; D000368:Aged; D002277:Carcinoma / Q000175:diagnosis* / Q000235:genetics*; D002291:Carcinoma, Papillary; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003124:Colorimetry; D004252:DNA Mutational Analysis / Q000379:methods*; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016612:Paraffin Embedding; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D000077273:Thyroid Cancer, Papillary; D013964:Thyroid Neoplasms / Q000175:diagnosis* / Q000235:genetics*; D016707:Tissue Fixation
D004273:DNA, Neoplasm; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf