METHOD FOR ESTIMATION OF BONE MARROW ASPIRATION QUALITY DURING MONITORING OF MINIMUM RESIDUAL DISEASE IN CASE OF MULTIPLE MYELOMA Russian patent published in 2017 - IPC G01N33/53 

FIELD: medicine.

SUBSTANCE: initially, bone marrow aspiration lysis is performed and after centrifugation the cell suspension is evaluated for the presence of nucleated cells and up to 3×10⁶ nucleated sample cells from the resulting precipitate are stained with monoclonal antibodies, the sample is incubated and analyzed. The resulting precipitate is stained using a six-colour panel of monoclonal antibodies: CD38 labelled with fluorescein isothiocyanate, CD138 labelled with phycoerythrin, CD45 labelled with allophycocyanine-Cy7, CD19 labelled with peridinine-chlorophyll-Cy5.5, CD56 labelled with phycoerythrin-Cy7, CD27, labelled with allophycocyanine. After incubation, cytometric analysis is performed and the entire population of plasma cells is isolated based on the expression of CD38, CD138 and CD45 markers. Expression levels of CD19, CD56, CD45 and CD27 markers on the plasma cells are estimated, and if two or more signs of anomalousness are present on the plasma cells, in particular the absence of CD19 expression, the absence of CD45 expression, the absence of CD27 expression or the presence of CD56 expression, the plasma cell is considered residual. And when such cells are detected in the test sample, it is analyzed for the presence of B-cell progenitors for CD45 and CD38 markers, and at their amount less than 0.33% of the whole population of nucleated cells of bone marrow suspension, the quality of bone marrow aspiration is assessed as unsatisfactory, that is, the sample is excessively diluted with peripheral blood, and in order to avoid a false negative result for the presence of minimal residual disease in multiple myeloma, it is recommended to repeat bone marrow puncture, and in case of presence of 0.33% and more of B-cell progenitors, the bone marrow sample quality is assessed as satisfactory and the proportion of residual plasma cells from all nucleated cells is indicated.

EFFECT: application of this method allows to assess the quality of bone marrow aspiration during monitoring of the minimal residual disease in multiple myeloma, including its study for the presence of a cell population by multicolour flow cytometry.

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