Minimal residual disease monitoring using multicolor flow cytometry and allele-specific polymerase chain reaction for the  MYD88  gene p.L265P mutation in patients with Waldenstrom’s macroglobulinemia

I. V. Galtseva; Гальцева И. В.; Anastasiya B. Vinnikova; Винникова Анастасия Борисовна; A. E. Grachev; Грачев А. Е.; A. A. Kulikov; Куликов А. А.; Yu. A. Tsoy; Цой Ю. А.; K. A. Nikiforova; Никифорова К. А.; N. M. Kapranov; Капранов Н. М.; S. M. Kulikov; Куликов С. М.; S. E. Starchenko; Старченко С. Э.; N. A. Severina; Северина Н. А.; E. E. Nikulina; Никулина Е. Е.; A. B. Sudarikov; Судариков А. Б.; E. O. Gribanova; Грибанова Е. О.; E. E. Zvonkov; Звонков Е. Е.; V. V. Troitskaya; Троицкая В. В.; E. N. Parovichnikova; Паровичникова Е. Н.

doi:10.17650/1818-8346-2026-21-2-53-63

Minimal residual disease monitoring using multicolor flow cytometry and allele-specific polymerase chain reaction for the MYD88 gene p.L265P mutation in patients with Waldenstrom’s macroglobulinemia

Authors: Galtseva I.V.¹, Vinnikova A.B.¹, Grachev A.E.¹, Kulikov A.A.², Tsoy Y.A.¹, Nikiforova K.A.¹, Kapranov N.M.¹, Kulikov S.M.¹, Starchenko S.E.¹, Severina N.A.¹, Nikulina E.E.¹, Sudarikov A.B.¹, Gribanova E.O.¹, Zvonkov E.E.¹, Troitskaya V.V.¹, Parovichnikova E.N.¹
Affiliations:
1. National Medical Research Center for Hematology, Ministry of Health of Russia
2. Lomonosov Moscow State University
Issue: Vol 21, No 2 (2026)
Pages: 53-63
Section: NEW DIRECTIONS, DIAGNOSTIC OPPORTUNITIES, AND TREATMENT ADVANCES
Published: 25.06.2026
URL: https://oncohematology.abvpress.ru/ongm/article/view/1104
DOI: https://doi.org/10.17650/1818-8346-2026-21-2-53-63
ID: 1104

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Abstract

Background. Waldenstrom’s macroglobulinemia (WM) is a rare indolent lymphoma characterized by bone marrow infiltration by lymphoplasmacytic cells and monoclonal immunoglobulin (Ig) M secretion. For primary diagnosis and assessment of remission depth, allele-specific polymerase chain reaction (AS-PCR) is also used to quantitatively assess the p.L265P mutation of the MYD88 gene with a sensitivity of 0.1 %. The treatment response is assessed according to the international NCCN (National Comprehensive Cancer Network) criteria, which are based on the regression of clinical symptoms and a decrease in serum monoclonal IgM concentration. However, this approach does not always reflect complete elimination of the tumor clone. Since the serum IgM concentration, determined by electrophoresis, can be maintained even in the absence of tumor cells in the bone marrow (up to 6 months), it is possible to use methods based on tumor clone characteristics, such as multicolor flow cytometry (MFC). The usefulness of minimal residual disease (MRD) monitoring using the MFC method, which is the standard for assessing remission depth in many other hematological malignancies, in WM has not been clearly determined.

Aim. To compare the MRD-positive / negative status by MFC with the presence / absence of MYD88 gene p.L265P mutation by AS-PCR and clinical and laboratory characteristics of WM patients.

Materials and methods. The study included 70 patients with WM who received treatment at the National Medical Research Center for Hematology between 2017 and 2025. The diagnosis was established in accordance with international criteria (2-IWWM (2^nd International Workshop on Waldenstrom’s Macroglobulinemia) 2002, NCCN 2025) and Russian clinical guidelines. To assess the remission depth, 13-color MFC and AS-PCR to detect MYD88 gene p.L265P mutation in bone marrow aspirate were used.

Results. Achieving MRD-negative status was statistically significantly more often associated with a lower baseline tumor burden (B-cell infiltration, IgM and β2-microglobulin levels) and higher hemoglobin and platelet counts. Regression of clinical manifestations (hepatosplenomegaly, B-symptoms) was not a clear predictor of achieving MRD-negative status. The study results confirmed the prognostic value of MRD monitoring by MFC for assessing the progression probability. The presence of MYD88 gene p.L265P mutation has not been confirmed as an unfavorable prognostic factor and continues to be studied.

Conclusion. MFC is a universal method for both primary diagnosis and MRD monitoring in WM. Comprehensive MRD monitoring using MFC and AS-PCR for MYD88 gene mutation has prognostic value in WM. Achieving a double negative status (MFC^– / MYD88^–) is an objective marker of maximal complete remission.

Keywords

Waldenstrom’s macroglobulinemia, immunophenotyping, minimal residual disease, molecular genetic testing, p.L265P mutation of the MYD88 gene, allele-specific polymerase chain reaction

References

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