МІНІМАЛЬНА ЗАЛИШКОВА ХВОРОБА ПРИ ГОСТРІЙ ЛІМФОБЛАСТНІЙ ЛЕЙКЕМІЇ: МЕТОДИ ВИЯВЛЕННЯ ТА КІЛЬКІСНОГО ВИЗНАЧЕННЯ
DOI:
https://doi.org/10.15407/exp-oncology.2026.02.098Ключові слова:
MRD, flow cytometry, molecular techniqueАнотація
дентифікація мінімальної залишкової хвороби (МЗХ) зазнала значного прогресу завдяки розвитку технологій, застосованих у цій галузі. Якщо спочатку вона базувалася виключно на морфології, згодом методологія визначення МЗХ зазнала докорінних змін та включила каріотипування, цитогенетику, проточну цитометрію та інші чутливі методи. В огляді обговорено ключові технічні аспекти успішного виявлення МЗХ i проаналізовано кореляцію між результатами, отриманими за допомогою проточної цитометрії та молекулярно-генетичних методів.
Посилання
Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revisionof the World Health Organization classification of lymphoidneoplasms. Blood. 2016;127(20):2375-2390. https://doi.org/10.1182/blood-2016-01-643569
Coustan-Smith E, Mullighan CG, Onciu M, et al. Early T-cell precursorleukaemia: a subtype of very high-risk acute lymphoblastic leukaemia. Lancet Oncol. 2009;10(2):147-156. https://doi.org/10.1016/S1470-2045(08)70314-0
Gaynon PS, Trigg ME, Heerema NA, et al. Children’s Cancer Group trials in childhoodacute lymphoblastic leukemia: 1983–1995. Leukemia. 2000;14(12):2223-2233. https://doi.org/10.1038/sj.leu.2401939
Schultz KR, PullenDJ, Sather HN, et al. Risk- and response-based classification of childhoodB-precursor acute lymphoblastic leukemia: a combinedanalysis of prognostic markers from the PediatricOncology Group (POG) and Children’sCancer Group (CCG). Blood. 2006;109(3):926-935. https://doi.org/10.1182/blood-2006-01-024729
Hunger SP, Loh ML, WhitlockJA, et al. COG Acute Lymphoblastic Leukemia Committee. Children’s OncologyGroup’s 2013 blueprintfor research: Acute lymphoblastic leukemia. Pediatr Blood Cancer. 2013;60:957-963. https://doi.org/10.1002/pbc.24420
Goldberg JM, Silverman LB, Levy DE, et al. Childhood T-cell acute lymphoblastic leukemia: the Dana-Farber Cancer Institute acute lymphoblastic leukemia consortium experience. J Clin Oncol. 2003;21(19):3616-3622. https://doi.org/10.1200/JCO.2003.10.116
Wood BL, Winter SS, DunsmoreKP, et al. T-lymphoblastic Leukemia (T-ALL) shows excellent outcome, lack of significance of the early thymic precursor(ETP) immunophenotype, and validation of the prognostic value of end-induction minimalresidual disease(MRD) in Children’s OncologyGroup (COG) study AALL0434. Blood.2014;124(21):1.https://doi.org/10.1182/blood.V124.21.1.1
Smith M, Arthur D, Camitta B, et al. Uniformapproach to risk classification and treatment assignment for children withacute lymphoblastic leukemia.J ClinOncol. 1996;14(1):18-24. https://doi.org/10.1200/JCO.1996.14.1.18
Pullen J, Shuster JJ, Link M, et al. Significance of commonly used prognostic factors differs for childrenwith T cell acute lymphocyticleukemia (ALL), as comparedto those with B-precursor ALL. A Pediatric Oncology Group (POG)study. Leukemia. 1999;13(11):1696-1707. https://doi.org/10.1038/sj.leu.2401555
Rambaldi A, Borleri G, Dotti G, et al. Innovativetwo-step negative selectionof granulocyte colony-stimulating factor–mobilized circulating progenitor cells: Adequacy for autologous and allogeneic transplantation. Blood.1998;91(6):2189-2196.https://doi.org/10.1182/blood.V91.6.2189
Gaipa G, BassoG, Biondi A, et al. Detectionof minimal residualdisease in pediatric acute lymphoblastic leukemia. Cytometry B Clin Cytom. 2013;84(6):359-369. https://doi.org/10.1002/cyto.b.21101
Mancini M, Cedrone M, Diverio D, et al. Use of dual-color interphase FISH for the detectionof inv(16) in acute myeloid leukemiaat diagnosis, relapse and during follow-up: a study of 23 patients. Leukemia.2000;14(3):364-368. https://doi.org/10.1038/sj.leu.2401678
Bielorai B, Golan H, Trakhtenbrot L, et al. Combined analysis of morphology and fluorescence in situ hybridiza- tion in follow-upof minimal residualdisease in a child with Philadelphia-positive acute lymphoblastic leukemia. Cancer GenetCytogenet. 2002;138(1):64-68. https://doi.org/10.1016/S0165-4608(02)00578-2
Coustan-Smith E, Behm FG, Sanchez J, et al. Immunological detection of minimalresidual diseasein children with acute lymphoblastic leukaemia. Lancet. 1998;351(9102):550-554.https://doi.org/10.1016/S0140-6736(97)10295-1
Farahat N, Morilla A, Owusu-Ankomah K, et al. Detection of minimalresidual diseasein B-lineage acute lym- phoblastic leukaemia by quantitative flow cytometry. Br J Haematol. 1998;101(1):158-164. https://doi.org/10.1046/j.1365-2141.1998.00675.x
Coustan-Smith E, Sancho J, Hancock ML, et al. Clinicalimportance of minimalresidual diseasein childhood acute lymphoblasticleukemia. Blood. 2000;96(8):2691-2696. https://doi.org/10.1182/blood.V96.8.2691
San Miguel JF, Vidriales MB, López-Berges C, et al. Early immunophenotypical evaluation of minimalresidual diseasein acute myeloidleukemia identifies different patientrisk groups and may contribute to postinduction treatment stratification. Blood. 2001;98(6):1746-1751. https://doi.org/10.1182/blood.V98.6.1746.
Dworzak MN, Fröschl G, PrintzD, et al.; AustrianBerlin-Frankfurt-Münster Study Group. Prognostic significance and modalities of flow cytometric minimal residualdisease detection in childhoodacute lymphoblastic leukemia. Blood. 2002;99(6):1952-1958. https://doi.org/10.1182/blood.V99.6.1952
Coustan-Smith E, Sancho J, Behm FG, et al. Prognostic importance of measuring early clearance of leukemiccells by flow cytometryin childhood acute lymphoblastic leukemia. Blood. 2002;100(1):52-58. https://do.org/10.1182/blood-2002-01-0006
Coustan-Smith E, Sancho J, Hancock ML, et al. Use of peripheral bloodinstead of bone marrow to monitor residual diseasein children with acute lymphoblastic leukemia. Blood.2002;100(7):2399-2402. https://doi.org/10.1182/blood-2002-04-1130
Neale GA, Coustan-Smith E, Pan Q, et al. Tandem application of flow cytometryand polymerase chain reaction for comprehensive detection of minimalresidual diseasein childhood acute lymphoblastic leukemia. Leukemia.1999;13(8):1221-1226. https://doi.org/10.1038/sj.leu.2401459
Oelschlägel U, Nowak R, Schaub A, et al. Shift ofaberrant antigen expression atrelapse orattreatment failure in acute leuke- mia. Cytometry.2000;42(4):247-253. https://doi.org/10.1002/1097-0320(20000815)42:4<247::AID-CYTO5>3.0.CO;2-V
Baer MR, Stewart CC, Dodge RK, et al. High frequencyof immunophenotype changes in acute myeloid leuke-mia at relapse:implications for residual diseasedetection (Cancer and LeukemiaGroup B Study 8361). Blood. 2001;97(11):3574-3580. https://doi.org/10.1182/blood.V97.11.3574
Porwit-MacDonald A, Björklund E, Lucio P, et al. BIOMED-1 concerted action report: Flow cytometric charac- terization of CD7+ cell subsets in normal bone marrow as a basis for the diagnosisand follow-up of T cell acute lymphoblastic leukemia (T-ALL). Leukemia.2000;14(5):816-825.https://doi.org/10.1038/sj.leu.2401741
Campana D, Coustan-Smith E. Advances in the immunological monitoring of childhood acute lymphoblastic leu- kaemia. BestPract Res Clin Haematol.2002;15(1):1-19. https://doi.org/10.1053/beha.2002.0182
Dworzak MN, Fritsch G, Fleischer C, et al. Comparative phenotype mapping of normal vs. malignant pediatric B-lymphopoiesis unveils leukemia-associated aberrations. Exp Hematol.1998;26(4):305-313. PMID: 9546313
Ciudad J, San Miguel JF, López-Berges MC, et al. Detectionof abnormalities in B-cell differentiation pattern is a useful tool to predict relapse in precursor-B-ALL. Br J Haematol. 1999;104(4):695-705. https://doi.org/10.1046/j.1365-2141.1999.01236.x
Fuda F, Chen W. Minimal/measurable residual disease detectionin acute leukemiasby multiparameter flow cyto- metry. Curr HematolMalig Rep. 2018;13(6):455-466. https://doi.org/10.1007/s11899-018-0479-1
Lavabre-Bertrand T, Janossy G, Ivory K, et al. Leukemia-associated changes identified by quantitative flow cyto- metry: I. CD10 expression. Cytometry. 1994;18(4):209-217. https://doi.org/10.1002/cyto.990180404
Flores-Montero J, Sanoja-Flores L, Paiva B, et al. Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma. Leukemia. 2017;31(10):2094-2103. https://doi.org/10.1038/leu.2017.29
Theunissen P, Mejstrikova E, Sedek L, et al. Standardized flow cytometryfor highly sensitiveMRD measurements in B-cell acutelymphoblastic leukemia. Blood. 2017;129(3):347-357. https://doi.org/10.1182/blood-2016-07-726307
Della Starza I, ChiarettiS, De Propris MS, et al. Minimal residual diseasein acute lymphoblastic leukemia: Techni- cal andclinical advances. Front Oncol.2019;9:726. https://doi.org/10.3389/fonc.2019.00726
Theunissen P, Mejstrikova E, Sedek L, et al. Standardized flow cytometryfor highly sensitiveMRD measurements in B-cell acutelymphoblastic leukemia. Blood. 2017;129(3):347-357. https://doi.org/10.1182/blood-2016-07-726307
Liu Z, Li Y, Shi C. Monitoring minimal/measurable residual diseasein B-cell acute lymphoblastic leukemia by flow cytometryduring targeted therapy. Int J Hematol. 2021;113(3):337-343. https://doi.org/10.1007/s12185-021-03085-y
Kruse A, Abdel-Azim N, Kim HN, et al. Minimalresidual disease detectionin acute lymphoblastic leukemia. Int JMol Sci. 2020;21(3):1054. https://doi.org/10.3390/ijms21031054
Szczepański T, Beishuizen A, Pongers-Willemse MJ, etal. Cross-lineage T cell receptor gene rearrangements occurin more than ninetypercent of childhood precursor-Bacute lymphoblastic leukemias:Alternative PCR targets for detectionof minimal residual disease. Leukemia.1999;13(2):196-205. https://doi.org/10.1038/sj.leu.2401277
Langerak AW, Groenen PJTA, Brüggemann M, et al. EuroClonality/BIOMED-2 guidelines for interpretation and reporting of Ig/TCR clonality testing in suspectedlymphoproliferations. Leukemia.2012;26(10):2159-2171. https://doi.org/10.1038/leu.2012.246
Langerak AW, Szczepański T, van der Burg M, et al. Heteroduplex PCR analysisof rearranged T cell receptor genes for clonality assessment in suspect T cell proliferations. Leukemia.1997;11(12):2192-2199. https://doi.org/10.1038/sj.leu.2400887
Germano G, del Giudice L, PalatronS, et al. Clonality profile in relapsedprecursor-B-ALL children by GeneScanand sequencing analyses. Consequences on minimalresidual diseasemonitoring. Leukemia.2003;17(8):1573-1582. https://doi.org/10.1038/sj.leu.2403008
Verhagen O, Willemse MJ, Breunis WB, et al. Application of germline IGH probes in real-timequantitative PCR for the detection of minimal residualdisease in acute lymphoblastic leukemia. Leukemia.2000;14(8):1426-1435. https://doi.org/10.1038/sj.leu.2401801
van der Velden VH, CazzanigaG, Schrauder A, et al.; EuropeanStudy Group on MRD detectionin ALL (ESG-MRD-ALL). Analysis of minimal residual disease by Ig/TCR gene rearrangements: guidelines for interpretation of real-timequantitative PCR data. Leukemia. 2007;21(4):604-611. https://doi.org/10.1038/sj.leu.2404586
van der Velden VH, NoordijkR, Brussee M, et al. Minimalresidual diseasediagnostics in acute lymphoblastic leukaemia: Impact of primer characteristics and size of junctional regions. Br J Haematol. 2014;164(3):451-453. https://doi.org/10.1111/bjh.12621
Szczepański T, Velden VHJ van der, Raff T, et al. Comparative analysis of T-cell receptor gene rearrangements at diagnosis and relapse of T-cell acute lymphoblastic leukemia (T-ALL) shows high stabilityof clonal markers for monitoring of minimalresidual diseaseand reveals the occurrence of second T-ALL. Leukemia.2003;17(11):2149-2156. https://doi.org/10.1038/sj.leu.2403081
Szczepanski T, Willemse MJ, Brinkhof B, et al. Comparative analysis of Ig and TCR gene rearrangements at diagno- sis and at relapse of childhoodprecursor-B–ALL provides improved strategies for selection of stable PCR targetsfor monitoring of minimal residual disease. Blood.2002;99(7):2315-2323. https://doi.org/10.1182/blood.v99.7.2315
Pui CH, RellingMV, Downing JR. Acute lymphoblastic leukemia. N Engl J Med. 2004;350(15):1535-1548. https://doi.org/10.1056/NEJMra023001
Downing JR, Shannon KM. Acute leukemia:A pediatric perspective. Cancer Cell. 2002;2(6):437-445. https://doi.org/10.1016/S1535-6108(02)00211-8
Ajuba IC, Madu AJ, Okocha C, et al. Frequencyand clinical impact of ETV6/RUNX1, AF4-MLL,and BCR/ABL fusion genes on features of acute lymphoblastic leukemia at presentation. Niger J Clin Pract. 2016;19(2):237-241. https://doi.org/10.4103/1119-3077.164351
Wu D, Sherwood A, Fromm JR, et al. High-throughput sequencing detects minimal residual disease in acute T lymphoblastic leukemia. Sci Transl Med. 2012;4(134):134ra63. https://doi.org/10.1126/scitranslmed.3003656
Wu D, Emerson RO, Sherwood A, et al. Detection of minimalresidual diseasein B lymphoblastic leukemia by high-throughput sequencing of IGH. Clin Cancer Res. 2014;20(17):4540-4548. https://doi.org/10.1158/1078-0432.CCR-13-3231
Wood B, Wu D, Crossley B, et al. Measurable residual diseasedetection by high-throughput sequencing improves risk stratificationfor pediatric B-ALL. Blood. 2018;131(12):1350-1359. https://doi.org/10.1182/blood-2017-09-806521
Wood BL, Wu D, Kirsch IM, et al. Residualdisease monitoringby high throughputsequencing provides risk stratifi- cation in childhood B-ALL and identifies a novel subset of patientshaving poor outcome. Blood.2016;128(22):1086. https://doi.org/10.1182/blood.V128.22.1086.1086
Pulsipher MA, Carlson C, LangholzB, et al. IgH-V(D)JNGS-MRD measurement pre-and early post-allotrans- plant defines very low-and very high-riskALL patients. Blood. 2015;125(22):3501-3508. https://doi.org/10.1182/blood-2014-12-615757
Kotrova M, Muzikova K, Mejstrikova E, et al. The predictive strength of next-generation sequencing MRD detec-tion for relapse compared with current methods in childhood ALL. Blood. 2015;126(8):1045-1047. https://doi.org/10.1182/blood-2015-07-655159
Huggett JF, Whale A. Digital PCR as a novel technology and its potential implications for molecular diagnostics. Clin Chem. 2013;59(12):1691-1693. https://doi.org/10.1373/clinchem.2013.214742
Huggett JF, Cowen S, Foy CA.Considerations for digital PCR as an accuratemolecular diagnostic tool. Clin Chem. 2015;61(1):79-88. https://doi.org/10.1373/clinchem.2013.214742
Reinert T, Schøler LV, Thomsen R, et al. Analysis of circulating tumour DNA tomonitor disease burden following colorectalcancer surgery. Gut. 2016;65(4):625-634. https://doi.org/10.1136/gutjnl-2014-308859
Waterhouse M, Follo M, PfeiferD, et al. Sensitive and accuratequantification of JAK2 V617F mutation in chronic myeloproliferative neoplasms by droplet digitalPCR. Ann Hematol. 2016;95(5):739-744. https://doi.org/10.1007/s00277-016-2623-0
Della Starza I, De Novi LA, Elia L, et al. Optimizing molecular minimalresidual diseaseanalysis in adult acute lymphoblastic leukemia. Cancers.2023;15(2):374. https://doi.org/10.3390/cancers15020374
Pecoraro S, BerbenG, Burns M, et al. Overview and recommendations for the application of digital PCR.Lux- embourg: Publications Office of the EuropeanUnion, ISBN 978-92-76-00180-5, Article115736. https://doi.org/10.2760/192883
van der Velden VH, CazzanigaG, Schrauder A, et al.; EuropeanStudy Group on MRD detectionin ALL (ESG-MRD-ALL). Analysis of minimal residual disease by Ig/TCR gene rearrangements: guidelines for interpretation of real-timequantitative PCR data. Leukemia. 2007;21:604-611. https://doi.org/10.1038/sj.leu.2404586
Fronkova E, Muzikova K, Mejstrikova E, et al. B-cell reconstitution after allogeneic SCT impairs minimalresidual diseasemonitoring in childrenwith ALL. Bone Marrow Transplant. 2008;42(3):187-196. https://doi.org/10.1038/bmt.2008
van Dongen JJ, van der Velden VH, Brüggemann M, et al. Minimal residual disease diagnostics in acute lympho-blastic leukemia: need for sensitive, fast, and standardized technologies. Blood. 2015;125(26):3996-4009. https://doi.org/10.1182/blood-2015-03-580027
Kotrova M, van der Velden VHJ, van DongenJJM, et al. Next-generation sequencing indicates false-positive MRD results and better predictsprognosis after SCT in patients with childhoodALL. Bone Marrow Transplant. 2017;52(7):962-968. https://doi.org/10.1038/bmt.2017.16
Kotrova M, Muzikova K, Mejstrikova E, et al. The predictive strength of next-generation sequencing MRD detec-tion for relapse compared with current methods in childhood ALL. Blood. 2015;126(8):1045-1047. https://doi.org/10.1182/blood-2015-07-655159
Eckert C, Flohr T, Köhler R, et al. Very early/early relapses of acute lymphoblastic leukemia show unexpected changes of clonal markers and high heterogeneity in responseto initial and relapse treatment. Leukemia.2011;25(8):1305-1313.https://doi.org/10.1038/leu.2011.89
Ladetto M, Brüggemann M, MonitilloL, et al. Next-generation sequencing and real-time quantitative PCR for minimal residual diseasedetection in B-cell disorders. Leukemia.2014;28(6):1299-1307. https://doi.org/10.1038/leu.2013.375
Submitted: September 10, 2025
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