High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories
Background: Monitoring of measurable residual disease (MRD) requires highly sensitive flow cytometry protocols to provide an accurate prediction of shorter progression-free survival. High assay sensitivity generally requires rapid processing to avoid cell loss from small bone marrow sample volumes,...
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MDPI AG
2024-12-01
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| Series: | Diseases |
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| Online Access: | https://www.mdpi.com/2079-9721/12/12/338 |
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| author | María Beatriz Álvarez Flores María Sopeña Corvinos Raquel Guillén Santos Fernando Cava Valenciano |
| author_facet | María Beatriz Álvarez Flores María Sopeña Corvinos Raquel Guillén Santos Fernando Cava Valenciano |
| author_sort | María Beatriz Álvarez Flores |
| collection | DOAJ |
| description | Background: Monitoring of measurable residual disease (MRD) requires highly sensitive flow cytometry protocols to provide an accurate prediction of shorter progression-free survival. High assay sensitivity generally requires rapid processing to avoid cell loss from small bone marrow sample volumes, but this requirement conflicts with the need in most clinical cytometry laboratories for long processing and acquisition times, especially when multiple MRD studies coincide on the same day. Methods: The proposed protocol was applied to 226 human bone marrow and 45 peripheral blood samples submitted for the study of MRD or the detection of rare cells. Samples were processed within 24 h of extraction and acquired with an eight-color flow cytometer. Results: The FACSLyse-Bulk protocol allows for the labelling of millions of cells in under 90 min in small sample volumes without affecting the FSC/SSC pattern or antigen expression, and it also allows antigens to be fixed to the membrane, thus avoiding the capping phenomenon. Conclusions: The proposed protocol would allow clinical flow cytometry laboratories to perform MRD studies in house and easily achieve a limit of detection and limit of quantification <0.001%, thus avoiding the need to outsource analysis to specialized cytometry laboratories. |
| format | Article |
| id | doaj-art-8027f0fb1f9b4f5b87fd3d7be07d1f53 |
| institution | Kabale University |
| issn | 2079-9721 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diseases |
| spelling | doaj-art-8027f0fb1f9b4f5b87fd3d7be07d1f532024-12-27T14:21:24ZengMDPI AGDiseases2079-97212024-12-01121233810.3390/diseases12120338High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical LaboratoriesMaría Beatriz Álvarez Flores0María Sopeña Corvinos1Raquel Guillén Santos2Fernando Cava Valenciano3Centro Nacional de Investigaciones Cardiovasculares Carlos III, 28029 Madrid, SpainURSalud Laboratory, Hospital Universitario Infanta Sofia, 28702 Madrid, SpainURSalud Laboratory, Hospital Universitario Infanta Sofia, 28702 Madrid, SpainURSalud Laboratory, Hospital Universitario Infanta Sofia, 28702 Madrid, SpainBackground: Monitoring of measurable residual disease (MRD) requires highly sensitive flow cytometry protocols to provide an accurate prediction of shorter progression-free survival. High assay sensitivity generally requires rapid processing to avoid cell loss from small bone marrow sample volumes, but this requirement conflicts with the need in most clinical cytometry laboratories for long processing and acquisition times, especially when multiple MRD studies coincide on the same day. Methods: The proposed protocol was applied to 226 human bone marrow and 45 peripheral blood samples submitted for the study of MRD or the detection of rare cells. Samples were processed within 24 h of extraction and acquired with an eight-color flow cytometer. Results: The FACSLyse-Bulk protocol allows for the labelling of millions of cells in under 90 min in small sample volumes without affecting the FSC/SSC pattern or antigen expression, and it also allows antigens to be fixed to the membrane, thus avoiding the capping phenomenon. Conclusions: The proposed protocol would allow clinical flow cytometry laboratories to perform MRD studies in house and easily achieve a limit of detection and limit of quantification <0.001%, thus avoiding the need to outsource analysis to specialized cytometry laboratories.https://www.mdpi.com/2079-9721/12/12/338measurable residual diseasehematologyleukemialymphomaflow cytometrylimit of detection |
| spellingShingle | María Beatriz Álvarez Flores María Sopeña Corvinos Raquel Guillén Santos Fernando Cava Valenciano High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories Diseases measurable residual disease hematology leukemia lymphoma flow cytometry limit of detection |
| title | High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories |
| title_full | High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories |
| title_fullStr | High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories |
| title_full_unstemmed | High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories |
| title_short | High-Sensitivity Flow Cytometry for the Reliable Detection of Measurable Residual Disease in Hematological Malignancies in Clinical Laboratories |
| title_sort | high sensitivity flow cytometry for the reliable detection of measurable residual disease in hematological malignancies in clinical laboratories |
| topic | measurable residual disease hematology leukemia lymphoma flow cytometry limit of detection |
| url | https://www.mdpi.com/2079-9721/12/12/338 |
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