High-Resolution Melting Is a Sensitive, Cost-Effective, Time-Saving Technique for BRAF V600E Detection in Thyroid FNAB Washing Liquid: A Prospective Cohort Study

Abstract

Objective: The diagnostic accuracy of thyroid fine needle aspiration biopsy (FNAB) can be improved by the combination of cytological and molecular analysis. In this study, washing liquids of FNAB (wFNAB) were tested for the BRAF V600E mutation, using the sensitive and cost-effective technique called high-resolution melting (HRM). The aim was to demonstrate the feasibility of BRAF analysis in wFNAB and its diagnostic utility, combined with cytology. Design: Prospective cohort study. Methods: 481 patients, corresponding to 648 FNAB samples, were subjected to both cytological (on cells smeared onto a glass slide) and molecular analysis (on fluids obtained washing the FNAB needle with 1 ml of saline) of the same aspiration. BRAF V600E analysis was performed by HRM after methodological validation for application to wFNAB (technique sensitivity: 5.4%). Results: The cytological results of the FNAB were: 136 (21%) nondiagnostic (THY1); 415 (64%) benign (THY2); 80 (12.4%) indeterminate (THY3); 9 (1.4%) suspicious for malignancy (THY4); 8 (1.2%) diagnostic of malignancy (THY5). The BRAF V600E mutation was found in 5 THY2, 2 THY3, 6 THY4 and 6 THY5 samples. Papillary carcinoma diagnosis was histologically confirmed in all BRAF+ thyroidectomized patients. BRAF combined with cytology improved the diagnostic value compared to cytology alone in a subgroup of 74 operated patients. Conclusions: HRM was demonstrated to be a feasible method for BRAF analysis in wFNAB. Thanks to its sensitivity and cost-effectiveness, it might be routinely used on a large scale in clinical practice. In perspective, standby wFNAB samples could be analyzed a posteriori in case of indeterminate cytology and/or suspicious findings on ultrasound.

BRAFV600E mutation test on fine‐needle aspiration specimens of thyroid nodules: Clinical correlations for 4600 patients

“…Third, the ratio of PTC to other thyroid cancers may also affect the sensitivity of BRAF V600E mutation evaluation since BRAF V600E mutation is absent in follicular thyroid carcinomas and occurs at a lower rate in follicular variants of PTC. 53 Although the BRAF V600E test would allow clinicians to optimize management to avoid surgery for benign nodules, thereby decreasing health costs and postsurgical complications, 54 nearly 20% of PTC cases could not be properly distinguished preoperatively by BRAF V600E mutation detection, even when combined with FNA. This is likely related to the complexity of the carcinogenesis of PTC.…”

High-resolution melting analysis for rapid and sensitive NOTCH1 screening in chronic lymphocytic leukemia

“…To the best of our knowledge, HRM analysis has already been applied for the diagnosis of tumors and genetic diseases. HRM analysis has been confirmed to a reliable, accurate, and rapid screening method for APC mutations in oral squamous cell carcinoma (44). BRCA1 gene screening using HRM The NOTCH1 mutation analysis method TAT is defined as the time spent from measuring DNA extraction to the time that the results are generated from the instrument; it excludes the time spent for DNA extraction.…”

The use of high-resolution melting techniques for mutation screening of diseases caused by trinucleotide repeats expansion, with emphasis on the <em> AR </em> gene

“…Sanger sequencing has been considered to be the gold standard for CAG repeat length analysis; however, it is more a laborious, time-consuming, and expensive procedure. While HRM analysis is a costeffective, less laborious, and rapid technique and requires only one-step real-time PCR compared with Sanger sequencing 17,18 and restriction fragment length polymorphism-Southern blot analysis. 19 In developing countries with limited molecular diagnostic facilities, HRM analysis can be more useful, particularly for research and population screening.…”

“…HRM analysis is a cost-effective, less laborious, and rapid technique and requires only one-step realtime PCR compared with Sanger sequencing 17,18 and restriction fragment length polymorphism-Southern blot analysis. 19 In developing countries with limited Melting point (celcius degree)…”