STANDARDIZATION OF THE METHOD OF MULTI-TARGET SINGLE-NUCLEOTIDE ELONGATION FOR MOLECULAR GENETIC STUDY OF CYTOLOGICAL MATERIAL IN THYROID NEOPLASM

DOI: https://doi.org/10.29296/24999490-2020-06-05

P.A. Fedorova, V.D. Nazarov, A.A. Musaelyan, S.V. Lapin, M.E. Boriskova, U.V. Farafonova, V.L. Emanuel Center for Molecular Medicine, Pavlov First Saint-Petersburg State Medical University, Lva Tolstogo str., 6–8, Saint Petersburg, 197022, Russian Federation E-mail: [email protected]

Introduction. Thyroid cancer (TC) is the most prevalent cancer among malignant neoplasms of the organs of the endocrine system. Cytological categories Bethesda III–V of TC are considered to be the most difficult for making a diagnosis. Molecular genetic tests can be a tool to complement routine cytopathological studies. The aim of the study. To develop a panel for detecting point mutations in the cytological material of thyroid tumors using the multi-target single-nucleotide elongation (MSE) method. Materials and methods. The studied group of patients included 52 cases with thyroid neoplasm. Patients were divided into subgroups under Bethesda category IV, V and VI. The first, second, and third subgroup consisted of 24, 7, and 21 patients correspondingly. Cytological material was obtained by fine-needle aspiration biopsy. Genetic testing was carried out using the MSE approach. There were created 3 panels included the most common mutations in thyroid cancer: mutations in the BRAF, KRAS, NRAS, HRAS genes. The obtained results of a postoperative histological examination included 11 samples of papillary thyroid cancer, 9 samples of the follicular variant of papillary cancer and 5 - follicular adenoma. Results. 39% of cytological samples showed mutations. The BRAFV600E mutation was revealed in 29% out of founding aberration, NRASQ61R mutation in 10% of those. To verify the method, there were used positive and negative control samples confirmed by PCR. In all control samples, the results were completely consistent with the molecular genetic study by the MSE method. Conclusion. MSE is a highly sensitive and promising method for the detection of mutations in cytological samples in TC patients
Keywords: 
multi-target single-base elongation (MSE), thyroid cancer, somatic mutation
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