Nacchio, Mariantonia (2020) Design and validation of a next generation sequencing custom panel for the pre-surgical risk stratification of indeterminate thyroid fine needle aspirations. [Tesi di dottorato]

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Item Type: Tesi di dottorato
Lingua: English
Title: Design and validation of a next generation sequencing custom panel for the pre-surgical risk stratification of indeterminate thyroid fine needle aspirations
Date: 12 March 2020
Number of Pages: 47
Institution: Università degli Studi di Napoli Federico II
Department: Sanità Pubblica
Dottorato: Sanità pubblica e medicina preventiva
Ciclo di dottorato: 32
Coordinatore del Corso di dottorato:
Troncone, GiancarloUNSPECIFIED
Date: 12 March 2020
Number of Pages: 47
Uncontrolled Keywords: Thyroid,NGS,BRAF
Settori scientifico-disciplinari del MIUR: Area 06 - Scienze mediche > MED/08 - Anatomia patologica
Date Deposited: 23 Mar 2020 10:48
Last Modified: 08 Nov 2021 14:34


Background: Thyroid carcinoma is the most common malignant neoplasm of the endocrine system, with a growing incidence peak in the world (1). Thyroid tumors generally consist of benign lesions that can affect one lobe or both; however, only a small fraction of them are classified as malignant (7-15%). The correct classification of thyroid tumors represents a crucial point for the stratification of patients with thyroid cancer in terms of prognosis and therapy. Currently, aspiration of the fine needle of the thyroid (FNA) is the most reliable and economic diagnostic tool to evaluate the morphological characteristics of the thyroid neoplasm, but very scarce nucleic acids are available to perform molecular tests. Methods: A commercially available test based on real-time PCR (RT-PCR) technology was tested on a series of indeterminate thyroid FNAs. In addition, a customized NGS gene panel was designed and tested on a different set of retrospective FNA samples. Results: the RT-PCR assay, which evaluates the genomic alterations of BRAF, N-H-KRAS, RET / PTC and PAX8 / PPARG, was chosen for subsequent clinical validation on a prospective series of n = 1172 thyroid FNA. In addition, 76 samples were tested to validate the gene panel. The custum gene panel allows to widen the reference reference range. In fact, with it it is possible to test 15 important genes for the stratification and evaluation of ROM in thyroid nodules. Conclusions: Our preliminary data show that 7 RT-qPCR genes and a customized NGS panel are feasible tests that should be implemented in routine diagnostics in order to avoid surgical treatment for low-risk nodules. The significant difference in post-test ROM between MT-pos and MT-neg FNA confirmed the high positive predictive value of BRAFV600E and BRAF type mutations compared to RAS-like genomic alterations. Furthermore, the preliminary results of the customized NGS panel have been satisfactory enough to expect its actual future adoption on our routine clinical samples.


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