Comparison of Immunohistochemistry and Direct Sequencing Methods for Identification of the BRAFV600E Mutation in Papillary Thyroid Carcinoma

Kim, Jong Kyu; Seong, Chan Yong; Bae, In Eui; Yi, Jin Wook; Yu, Hyeong Won; Kim, Su jin; Won, Jae Kyung; Chai, Young Jun; Choi, June Young; Lee, Kyu Eun

doi:10.1245/s10434-018-6460-3

Cited by 33 publications

(34 citation statements)

References 31 publications

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“…A higher mutation rate was detected by ddPCR. These results were consistent with previous studies that showed a BRAF V600E mutation rate of 29%‐90% . In the present study, the BRAF V600E mutation was only detected in PTC patients and not in patients with benign nodular thyroid disease, which was also consistent with previous studies …”

Section: Discussionsupporting

confidence: 94%

“…Thyroid cancer (TC) is the most common type of endocrine malignancy, and papillary thyroid carcinoma (PTC) accounts for the vast majority (90%) of thyroid malignancies . BRAF V600E has a T1799A point mutation in exon 15, resulting in the substitution of valine for glutamic acid at amino acid 600 (V600E), and it is the most common driver mutation of PTC.…”

Section: Introductionmentioning

confidence: 99%

“…Many techniques can be used for the detection of the BRAF V600E mutation, and the gold standard method for the molecular diagnosis of the BRAF V600E mutation is direct Sanger sequencing . However, Sanger sequencing identified only 7%‐20% of mutated alleles in a background of wild‐type alleles …”

Section: Introductionmentioning

confidence: 99%

“…4,5 Many techniques can be used for the detection of the BRAF V600E mutation, [6][7][8] and the gold standard method for the molecular diagnosis of the BRAF V600E mutation is direct Sanger sequencing. 2 However, Sanger sequencing identified only 7%-20% of mutated alleles in a background of wild-type alleles. 8,9 Droplet digital PCR (ddPCR) is a recently introduced technology that may facilitate the detection of rare point mutations in a background of wild-type alleles.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAF^V600E mutation in papillary thyroid carcinoma

Wang

Sun²,

Jing

et al. 2019

Clinical Laboratory Analysis

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BackgroundThe BRAF V600E mutation status is a useful diagnostic and prognostic marker for papillary thyroid carcinoma (PTC). Although it is a commonly used method, Sanger sequencing has several limitations in detecting the BRAF V600E mutation. The aim of this study was to evaluate the efficiency of droplet digital PCR (ddPCR) as an alternative method for the detection of the BRAF V600E mutation in PTC patients.MethodsSamples from a total of 120 patients with PTC and 30 patients with benign nodular thyroid disease who underwent thyroid surgery were collected. The BRAF V600E mutation status of the PTC patients was tested by Sanger sequencing and ddPCR.ResultsThe BRAF V600E mutation was detected in 67 samples (44.67%) by Sanger sequencing and 92 samples (61.33%) by ddPCR. The detection of the mutation by the two methods was inconsistent in twenty‐five samples (16.67%). The sensitivity and specificity of the ddPCR method were 100% and 69.88%, respectively, and the positive predictive and negative predictive values were 72.83% and 100%, respectively. The concordance rate between the two methods in detecting the BRAF V600E mutation was 83.33%. Neither Sanger sequencing nor ddPCR detected BRAF V600E in 30 patients with benign nodular thyroid disease. The 92 samples with the BRAF V600E mutation were detected by ddPCR at a fractional abundance from 0.28% to 45.40% as follows: ≥10% (59 samples, 64.13%), 5%‐10% (8 samples, 8.70%), and ≤5% (25 samples, 27.17%). The BRAF V600E mutation was detected in all 59 samples at a fractional abundance ≥10% and in four samples at a fractional abundance from 5% to 10%, and no BRAF V600E mutation was detected at a fractional abundance ≤5% by Sanger sequencing.ConclusionsddPCR was a reliable, highly sensitive alternative method for the detection of the BRAF V600E mutation in PTC patients.

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Section: Discussionsupporting

confidence: 94%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAF^V600E mutation in papillary thyroid carcinoma

Wang

Sun²,

Jing

et al. 2019

Clinical Laboratory Analysis

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“…Anti-BRAF V600E (clone VE1) antibody, a mouse monoclonal primary antibody, is used in the identi cation of the BRAF V600E mutant protein. Previous studies have veri ed the sensitivity and speci city of anti-BRAF V600E (clone VE1) antibody in melanoma [7] and papillary thyroid carcinoma [8] . However, few studies have assessed the reliability of anti-BRAF-V600E immunohistochemistry (IHC) in brain tumors.…”

Section: Introductionmentioning

confidence: 99%

The Immunoreactivity Patterns of BRAF VE1 and Its Diagnostic Value in Brain Tumors

Fan

Chen³

et al. 2020

Preprint

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Background: BRAF mutations have been detected in a high proportion of melanoma, papillary thyroid carcinoma, and various primary brain tumors. But the sensitivity and specificity of immunohistochemical detection of BRAF-V600E mutant protein were not evaluated in brain tumors. The aim of this study was to assess the utlity of BRAF-V600E IHC compared to molecular biology on a large series of brain tumors, in order to provide a useful reference for the use of BRAF-V600E IHC in clinical practice.Methods and results: We analyzed the BRAF-V600E immunoreactivity pattern and its expression profile by immunohistochemistry (IHC) in 122 patients diagnosed with different tumors of brain including gangliocytoma/ganglioma(GC/GG), pleomorphic xanthoastrocytomas(PXA), epithelioid glioblastoma(E-GBM), dysembryoplastic neuroepithelial tumour(DNT), pilocytic astrocytoma(PA), and papillary craniopharyngioma(p-CPG). VE1 immunostaining of 52 cases showed clear cytoplasmic diffuse positive pattern in majority tumor cells. 14 cases were presented with clear granular cytoplasmic positive pattern in single tumor cell or tumor cell cluster. 22 cases displayed equivocal positive with undefined location. 34 cases were negative. Including 81 immunopositive cases and 29 immunonegative cases were further confirmed by Real-time PCR. And 63 of 81 immunopositive cases were confirmed with BRAF-V600E mutation (77.8%), and all of 29 negative cases were confirmed to have wild-type BRAF (100%). Interestingly, only the cases showing clear immunoreactivity patterns (e.g cytoplasmic) with clean background had immunostaining results consistent with the molecular detection results, regardless of the number of positive cells (61/61,100%). However, samples with indeterminate immunoreactivity patterns were most likely to have false positive results (18/20, 90%). Conclusions: VE1 immunostaining could replace molecular detection to some extent, on the premise of mastering the key points in the interpretation of BRAF VE1 immunostaining: 1) As long as the positive signal was accurately located in the cytoplasm of tumor cells, the sample was considered to have BRAF V600E mutation, disregarding the number of positive cells; 2) Tissue samples that had no signal of BRAF VE1 expression with clear background could be confirmed with wild-type BRAF-V600E; 3) Some equivocal positive with uniform “coating” or nucleus positive cases were often considered as false-positive and usually required further molecular detection.

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VE1 immunohistochemistry is an adjunct tool for detection of BRAF^V600E mutation: Validation in thyroid cancer patients

Rashid

Tabassum

Khan

et al. 2020

Clinical Laboratory Analysis

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Papillary thyroid carcinoma (PTC) is the most common endocrine malignancy among other endocrine tumors, and BRAFV600E is a frequent genetic mutation occurring in the disease. Although different molecular techniques, most importantly sequencing has been widely recognized as a gold standard but molecular diagnosis remains an expensive, laborious, and time‐intensive process. Recently, immunohistochemistry (IHC) with anti‐BRAF V600E (VE1) antibody has increased practical utility and implemented clinically for the detection of BRAFV600E mutation. Therefore, the study aimed to evaluate diagnostic accuracy of VE1 IHC for detecting the BRAFV600E mutation frequency and clinical implementation in diagnostic laboratories. In this study, 72 formalin fixed paraffin‐embedded tissues (FFPE) were used to determine the BRAFV600E mutation status using IHC and Sanger sequencing. The mutation was found in 29% and 28% cases using IHC and Sanger sequencing, respectively. Furthermore, the results showed 100% sensitivity, 98.07% specificity, 95.2% positive predictive value, and 100% negative predictive value. Notably, significant associations were found between BRAFV600E status and tumor stage, tumor focality, and extrathyroidal extensions, respectively. VE1 IHC was found to be a highly sensitive, specific, and diagnostically accurate method in this cohort. Therefore, BRAFV600E detection through IHC has been considered as the best tailored technique for routine pathology laboratories.

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Comparison of Immunohistochemistry and Direct Sequencing Methods for Identification of the BRAFV600E Mutation in Papillary Thyroid Carcinoma

Abstract: IHC using the VE1 antibody is a reliable and highly sensitive method for detecting the BRAF mutation in classic PTC.

Cited by 33 publications

References 31 publications

Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAF^V600E mutation in papillary thyroid carcinoma

Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAF^V600E mutation in papillary thyroid carcinoma

The Immunoreactivity Patterns of BRAF VE1 and Its Diagnostic Value in Brain Tumors

VE1 immunohistochemistry is an adjunct tool for detection of BRAF^V600E mutation: Validation in thyroid cancer patients

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Comparison of Immunohistochemistry and Direct Sequencing Methods for Identification of the BRAFV600E Mutation in Papillary Thyroid Carcinoma

Abstract: IHC using the VE1 antibody is a reliable and highly sensitive method for detecting the BRAF mutation in classic PTC.

Cited by 33 publications

References 31 publications

Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAFV600E mutation in papillary thyroid carcinoma

Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAFV600E mutation in papillary thyroid carcinoma

The Immunoreactivity Patterns of BRAF VE1 and Its Diagnostic Value in Brain Tumors

VE1 immunohistochemistry is an adjunct tool for detection of BRAFV600E mutation: Validation in thyroid cancer patients

Contact Info

Product

Resources

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Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAF^V600E mutation in papillary thyroid carcinoma

Comparison of droplet digital PCR and direct Sanger sequencing for the detection of the BRAF^V600E mutation in papillary thyroid carcinoma

VE1 immunohistochemistry is an adjunct tool for detection of BRAF^V600E mutation: Validation in thyroid cancer patients