Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction
Abstract Background Recent guidelines for the management of thyroid nodules incorporate mutation testing as an adjunct for surgical decision-making, however current tests are costly with limited accuracy. Droplet digital PCR (ddPCR) is an ultrasensitive method of nucleic acid detection that is parti...
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SAGE Publishing
2018-09-01
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| Series: | Journal of Otolaryngology - Head and Neck Surgery |
| Online Access: | http://link.springer.com/article/10.1186/s40463-018-0299-2 |
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| author | Vincent L. Biron Ashlee Matkin Morris Kostiuk Jordana Williams David W. Cote Jeffrey Harris Hadi Seikaly Daniel A. O’Connell |
| author_facet | Vincent L. Biron Ashlee Matkin Morris Kostiuk Jordana Williams David W. Cote Jeffrey Harris Hadi Seikaly Daniel A. O’Connell |
| author_sort | Vincent L. Biron |
| collection | DOAJ |
| description | Abstract Background Recent guidelines for the management of thyroid nodules incorporate mutation testing as an adjunct for surgical decision-making, however current tests are costly with limited accuracy. Droplet digital PCR (ddPCR) is an ultrasensitive method of nucleic acid detection that is particularly useful for identifying gene mutations. This study aimed to assess the analytic and clinical validity of RAS and BRAF ddPCR mutational testing as a diagnostic tool for thyroid fine needle aspirate biopsy (FNAB). Methods Patients with thyroid nodules meeting indication for FNAB were prospectively enrolled from March 2015 to September 2017. In addition to clinical protocol, an additional FNAB was obtained for ddPCR. Optimized ddPCR probes were used to detect mutations including HRASG12 V, HRASQ61K, HRASQ61R, NRASQ61R, NRASQ61K and BRAFV600E. The diagnostic performance of BRAF and RAS mutations was assessed individually or in combination with Bethesda classification against final surgical pathology. Results A total of 208 patients underwent FNAB and mutational testing with the following Bethesda cytologic classification: 26.9% non-diagnostic, 55.2% benign, 5.3% FLUS/AUS, 2.9% FN/SPN, 2.4% SFM and 7.2% malignant. Adequate RNA was obtained from 91.3% (190) FNABs from which mutations were identified in 21.1% of HRAS, 11.5% of NRAS and 7.4% of BRAF. Malignant cytology or BRAFV600E was 100% specific for malignancy. Combining cytology with ddPCR BRAF600E mutations testing increased the sensitivity of Bethesda classification from 41.7 to 75%. Combined BRAFV600E and Bethesda results had a positive predictive value (PPV) of 100% and negative predictive value (NPV) of 89.7% for thyroid malignancy in our cohort. Conclusions DdPCR offers a novel and ultrasensitive method of detecting RAS and BRAF mutations from thyroid FNABs. BRAFV600E mutation testing by ddPCR may serve as a useful adjunct to increase sensitivity and specificity of thyroid FNAB. |
| format | Article |
| id | doaj-art-b8173b4b6e0a4cae9f25a89bfe581b4a |
| institution | Kabale University |
| issn | 1916-0216 |
| language | English |
| publishDate | 2018-09-01 |
| publisher | SAGE Publishing |
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| series | Journal of Otolaryngology - Head and Neck Surgery |
| spelling | doaj-art-b8173b4b6e0a4cae9f25a89bfe581b4a2025-02-03T00:22:57ZengSAGE PublishingJournal of Otolaryngology - Head and Neck Surgery1916-02162018-09-014711910.1186/s40463-018-0299-2Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reactionVincent L. Biron0Ashlee Matkin1Morris Kostiuk2Jordana Williams3David W. Cote4Jeffrey Harris5Hadi Seikaly6Daniel A. O’Connell7Division of Otolaryngology-Head and Neck Surgery, University of AlbertaAlberta Head and Neck Centre for Oncology and Reconstruction, Walter MacKenzie Health Sciences CentreAlberta Head and Neck Centre for Oncology and Reconstruction, Walter MacKenzie Health Sciences CentreOtolaryngology-Head and Neck Surgery Research Laboratory of Alberta, University of AlbertaDivision of Otolaryngology-Head and Neck Surgery, University of AlbertaDivision of Otolaryngology-Head and Neck Surgery, University of AlbertaDivision of Otolaryngology-Head and Neck Surgery, University of AlbertaDivision of Otolaryngology-Head and Neck Surgery, University of AlbertaAbstract Background Recent guidelines for the management of thyroid nodules incorporate mutation testing as an adjunct for surgical decision-making, however current tests are costly with limited accuracy. Droplet digital PCR (ddPCR) is an ultrasensitive method of nucleic acid detection that is particularly useful for identifying gene mutations. This study aimed to assess the analytic and clinical validity of RAS and BRAF ddPCR mutational testing as a diagnostic tool for thyroid fine needle aspirate biopsy (FNAB). Methods Patients with thyroid nodules meeting indication for FNAB were prospectively enrolled from March 2015 to September 2017. In addition to clinical protocol, an additional FNAB was obtained for ddPCR. Optimized ddPCR probes were used to detect mutations including HRASG12 V, HRASQ61K, HRASQ61R, NRASQ61R, NRASQ61K and BRAFV600E. The diagnostic performance of BRAF and RAS mutations was assessed individually or in combination with Bethesda classification against final surgical pathology. Results A total of 208 patients underwent FNAB and mutational testing with the following Bethesda cytologic classification: 26.9% non-diagnostic, 55.2% benign, 5.3% FLUS/AUS, 2.9% FN/SPN, 2.4% SFM and 7.2% malignant. Adequate RNA was obtained from 91.3% (190) FNABs from which mutations were identified in 21.1% of HRAS, 11.5% of NRAS and 7.4% of BRAF. Malignant cytology or BRAFV600E was 100% specific for malignancy. Combining cytology with ddPCR BRAF600E mutations testing increased the sensitivity of Bethesda classification from 41.7 to 75%. Combined BRAFV600E and Bethesda results had a positive predictive value (PPV) of 100% and negative predictive value (NPV) of 89.7% for thyroid malignancy in our cohort. Conclusions DdPCR offers a novel and ultrasensitive method of detecting RAS and BRAF mutations from thyroid FNABs. BRAFV600E mutation testing by ddPCR may serve as a useful adjunct to increase sensitivity and specificity of thyroid FNAB.http://link.springer.com/article/10.1186/s40463-018-0299-2 |
| spellingShingle | Vincent L. Biron Ashlee Matkin Morris Kostiuk Jordana Williams David W. Cote Jeffrey Harris Hadi Seikaly Daniel A. O’Connell Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction Journal of Otolaryngology - Head and Neck Surgery |
| title | Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction |
| title_full | Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction |
| title_fullStr | Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction |
| title_full_unstemmed | Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction |
| title_short | Analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction |
| title_sort | analytic and clinical validity of thyroid nodule mutational profiling using droplet digital polymerase chain reaction |
| url | http://link.springer.com/article/10.1186/s40463-018-0299-2 |
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