E.I. Antonova1, N.V. Firsova2, A.B. Achilov3, A.R. Zimnurov4, E.V. Balacuk5

1–5 Scientific Research Center for Fundamental and Applied Problems of Bioecology and Biotechnology
Ulyanovsk State Pedagogical University named after I.N. Ulyanov (Ulyanovsk, Russia)

1 antonov_67@mail.ru, 2 n-firsova@mail.ru, 3 a.achilow@inbox.ru, 4 zimnurov75@gmail.com, 5 balacyxa@mail.ru

Nodular melanoma is the second most common type of melanoma, characterized by rapid vertical growth, which makes early diagnosis difficult. The amelanotic variant mimics various forms of tumors, with a disproportionately high mortality rate associated with late diagnosis, which explains the 89% of diagnostic errors. The integration of highly sensitive diagnostic tests into clinical studies becomes essential when histological examinations fail to provide a definitive diagnosis. Multiplex mutation detection methods using xMAP technology allow for quick results by simultaneously analyzing multiple parallel reactions of different targets.

Purpose of the work is the development of a multiplex domestic protocol xMAP-ASPE-xTAG with simultaneous determination of gene mutations (BRAF, MITF, CDKN2A) in one analyte using the example of a nodular melanoma subtype and its variants – pigmented and amelanotic, for early diagnosis of the most aggressive melanoma subtype, the development of which takes from several weeks to several months.

It has been revealed that wtBRAF NM, to a greater extent than mtBRAF NM and ANM, exhibit a hereditary pattern of development, which is confirmed by the identified mutations in the CDKN2A and MITF-M genes, which predispose to the development of melanoma along the lowCSD pathway. MITF increases the expression of CDKN2A. BRAFV600E mutations support both decreased and increased levels of MITF transcription, promoting the proliferation and survival of melanoma cells. All ANM mtBRAF and mtBRAF NM samples show a lower correlation with inheritance and follow the lowCSD and highCSD pathways. Three samples with the lowest DNA concentration and mutant DNA wtBRAF NM, and the remaining samples are mtBRAF. The xMAP-ASPE-xTAG protocol parameters are determined based on the total sample area and formation area, total and mutant DNA concentration, optimal reagent concentration, and microsphere number.

The work is carried out as part of the development of a domestic diagnostic test system using the xMAP-ASPE-xTAG technology for early diagnosis and detection of predisposition to melanoma development.

Antonova E.I., Firsova N.V., Achilov A.B., Zimnurov A.R., Balacuk E.V. Molecular and genetic landscape of nodal melanoma. Technologies of Living Systems. 2025. V. 22. № 4. Р. 57-65. DOI: https://doi.org/10.18127/j20700997-202504-07 (In Russian).

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