Application of BRAF, NRAS, KRAS mutations as markers for the detection of papillary thyroid cancer from FNAB specimens by pyrosequencing analysis
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Seo-Jin Park
Abstract
Background: BRAF V600E, the most common BRAF gene mutation, is detected in approximately 50% of sporadic papillary thyroid carcinoma (PTC) and may be associated with triggering tumorigenesis of PTC. The aim of our study was to discover additional mutations to increase the diagnostic performance of molecular tests in screening for thyroid cancer from fine needle aspiration biopsy (FNAB) specimens.
Methods: DNA was extracted from 120 freshly obtained FNAB specimens selected according to cytopathology grades of the Bethesda system. A conventional BRAF V600E test was carried out with real-time PCR, and further mutation screening for BRAF mutations in codons 464, 466, 469, NRAS and KRAS codons 12/13 and 61 was done by pyrosequencing. Histopathology reports were reviewed for those who underwent thyroidectomy (n=83).
Results: The real-time PCR method detected 45 BRAF V600E- positive cases whereas pyrosequencing detected 30 cases. Additional BRAF (n=4), NRAS (n=11) and KRAS (n=3) mutations were detected in 17 cases (one overlapping BRAF and NRAS mutation). Among 11 NRAS-mutated cases, eight were confirmed as PTC and one as FVPTC on histopathology reports. Five PTC-confirmed cases with BRAF V600E mutation showed additional mutations, all of which were NRAS mutations.
Discussion: Despite the higher sensitivity of real-time PCR for detecting BRAFV600E mutations, pyrosequencing easily detected additional point mutations. NRAS mutations were the most prevalently identified additional mutations and were highly associated with malignancy. In conclusion, our findings demonstrate that additional mutations identified by pyrosequencing may help in the pre-operative process in determining the possibility of malignancy and further studies on the occurrence of simultaneous mutations of BRAF, KRAS and NRAS may be warranted.
References
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©2013 by Walter de Gruyter Berlin Boston
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Articles in the same Issue
- Letters to the Editors
- Clinical utility of serum tumor markers and cytokines in cervical cancer and neoplasia
- Establishing reference intervals for LDL subfractions in a Korean population using the Lipoprint LDL system
- Measurement imprecision of common urinary biochemical analytes on the Roche Cobas 6000 system
- A comparison between turbidimetric inhibition immunoassay and capillary electrophoresis in glycated hemoglobin (HbA1c) measurement
- Commutability: a peculiar property of calibration and control materials. Definition and evaluation
- Diagnostic sensitivity of a panel of tests to detect monoclonal protein in Korean multiple myeloma patients
- Commutability of proficiency testing (PT): status of the matrix-related bias in general clinical chemistry
- Masthead
- Masthead
- Editorial
- Why specifications for allowable glucose meter errors should include 100% of the data
- Reviews
- ABO blood group: old dogma, new perspectives
- Trace elements and bone health
- Mini Review
- The role of transcription factors in laboratory medicine
- Opinion Papers
- Nobelitis: a common disease among Nobel laureates?
- The syndrome of the “obsessive-compulsory scientist”: a new mental disorder?
- Can current analytical quality performance of UK clinical laboratories support evidence-based guidelines for diabetes and ischaemic heart disease? – A pilot study and a proposal
- Guidelines and Recommendations
- Survey of national guidelines, education and training on phlebotomy in 28 European countries: an original report by the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) working group for the preanalytical phase (WG-PA)
- General Clinical Chemistry and Laboratory Medicine
- Red cell indices: differentiation between β-thalassemia trait and iron deficiency anemia and application to sickle cell disease and sickle cell thalassemia
- Problems in determining thalassemia carrier status in a program for prevention and control of severe thalassemia syndromes: a lesson from Thailand
- An enzyme linked immunosorbent assay (ELISA) for the determination of the human haptoglobin phenotype
- Blood loss from laboratory diagnostic tests in children
- Hematocrit correction does not improve glucose monitor accuracy in the assessment of neonatal hypoglycemia
- Evaluation of a mobile clinical pathology laboratory developed for the home care of pediatric patients following transplantation of peripheral blood precursor cells
- Folic acid supplementation does not reduce intracellular homocysteine, and may disturb intracellular one-carbon metabolism
- Influence of spurious hemolysis on blood gas analysis
- Serum procalcitonin predicts development of acute kidney injury in patients with suspected infection
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- Nationwide multicenter study aimed at the establishment of common reference intervals for standardized clinical laboratory tests in Japan
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