A single tube 10‐color flow cytometry assay optimizes detection of minimal residual disease in chronic lymphocytic leukemia

Sartor, Mary; Gottlieb, David

doi:10.1002/cyto.b.21067

Cited by 26 publications

(15 citation statements)

References 17 publications

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“…The presence of MRD was reported as the percentage of pathological cells within the total leucocyte population. According to the literature, MRD was defined as positive when an accumulation of at least 20 clustered events displaying the above mentioned immunophenotypic characteristics was detected (Böttcher et al , ; Sartor & Gottlieb, ) with a sensitivity limit equal to 0·01%.…”

Section: Methodsmentioning

confidence: 99%

“…The proposed monoclonal antibody (MoAb) combinations, applicable to all sample types and therapeutic regimens, enable an accurate separation of CLL cells from the background, providing a high concordance with ASO IgH RQ‐PCR for the detection of CLL above 0·01% (10 −4 ). Moreover, instrument improvements and advances in FC methodology up to the use of 10 colour combinations (Sartor & Gottlieb, ) have reduced the number of required cells and allowed, at the same time, the analysis of a large number of events with an increase in sensitivity and accuracy.…”

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confidence: 99%

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Minimal residual disease monitoring in chronic lymphocytic leukaemia patients. A comparative analysis of flow cytometry and ASO IgH RQ‐PCR

et al. 2014

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SummaryMinimal residual disease (MRD) is becoming increasingly important in chronic lymphocytic leukaemia (CLL) as treatment strategies are progressively improving. The primary objective of this study was to compare the applicability of three different flow cytometric approaches: basic 4-colour analysis, European Research Initiative in CLL (ERIC) consensus method and 8-colour analysis. Secondly, we investigated the sensitivity and specificity of flow cytometry (FC) compared to molecular analyses for MRD detection. A total of 462 CLL samples were evaluated by basic FC; in 143, ERIC consensus method was also performed and all three FC methodologies were applied in a subgroup of 10 cases. No discordance in defining MRD-positive/negative samples was observed between the FC methods; within positive samples, the ERIC consensus method and 8-colour analysis showed the most accurate results. MRD was analysed by FC and polymerase chain reaction (PCR) in 243 cases: concordant results were obtained in 199/243 samples (81Á9%); 42/243 were FCÀ/PCR+. Overall, the sensitivity and specificity of FC compared to PCR was 96Á5% and 77Á2%, respectively. Both FC and PCR proved suitable for the detection of MRD and prediction of progression-free survival, which was significantly reduced in MRD-positive patients, regardless of the methodology. These results offer the rationale for a strategy to monitor MRD in CLL patients.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Minimal residual disease monitoring in chronic lymphocytic leukaemia patients. A comparative analysis of flow cytometry and ASO IgH RQ‐PCR

et al. 2014

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“…Furthermore, many studies concentrate on one gender or on a smaller age range, or comprise only younger and elderly, but with a gap in the intermediate ages. While specific clinical aspect have been characterized , in‐depth immunophenotyping of healthy individuals is seldom performed. However, since leukocyte subset counts vary with age, gender, and ethnicity reference intervals for Europeans from healthy subjects are needed that are able to reflect these influences.…”

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confidence: 99%

Reference intervals for leukocyte subsets in adults: Results from a population‐based study using 10‐color flow cytometry

Melzer

Zachariae

Bocsi

et al. 2015

Cytometry Part B Clinical

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Background: Reference intervals for leukocyte subsets from peripheral blood are helpful for the understanding of disease states and therapy effects.Methods: We performed in-depth immunophenotyping for 608 healthy German adults from the Leipzig region from 40 to 79 years by 10-color flow cytometry (FCM) to gain reference information for various leukocyte subsets including subsets of granulocytes, monocytes and lymphocytes.Results: First, we derived gender-and age-specific reference intervals for males and females from 40 to 59 and from 60 to 79 years, respectively. Second, we further investigated the influence of gender and age on leukocyte counts. We found significantly higher cell counts for monocytes (P < 0.001) and NK cells (P < 0.001) in men, whereas women had higher counts for B cells (P < 0.001), Th cells (P < 0.001) and regulatory T cells (P 5 0.008). Furthermore, with increasing age, a decrease in Tc cells (about 8% within 5 years) and an increase in NK cells (<4% within 5 years) were observed.Conclusion: In future research, it should be investigated whether these are real ageing effects that can be confirmed in longitudinal studies. Furthermore, it is important to understand if the Tc cell count drop is functionally compensated by the increase of NK cells. V C 2015 International Clinical Cytometry Society

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“…Do badania 8-kolorowego wykorzystuje się mniejszą liczbę przeciwciał, zastosowanie jednej probówki pozwala ocenić większą liczbę komórek, zaś analiza jest prostsza i trwa krócej niż w badaniach 4-i 6-kolorowych. Zastosowanie 8-lub więcej kolorowego, 1-probówkowego protokołu pozwala na osiągnięcie wyższej czułości, sięgającej 10 -5 -10 -6 [31,33].…”

Section: Cytometria Przepływowaunclassified

Znaczenie oceny minimalnej choroby resztkowej u chorych na przewlekłą białaczkę limfocytową

2018

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Przewlekła białaczka limfocytowa (CLL) jest chorobą limfoproliferacyjną o zróżnicowanym przebiegu klinicznym wywodzącą się z dojrzałych limfocytów B. W około 60% przypadków ze względu na zaawansowanie choroby konieczne jest rozpoczęcie leczenia. Zastosowanie intensywnej immunochemioterapii pozwala uzyskać długotrwałe remisje u części pacjentów, niemniej mimo uzyskania całkowitych odpowiedzi dochodzi do wznowy choroby. Jej nawrót tłumaczy się obecnością rezydualnych komórek białaczkowych w szpiku kostnym, krwi obwodowej czy węzłach chłonnych, określanych mianem minimalnej choroby resztkowej (MRD). Zastosowanie cytometrii przepływowej (FLC) oraz metod molekularnych umożliwia wykrycie pozostałych komórek białaczkowych z czułością sięgającą co najmniej 10-5 , co pozwala na dokładniejsze określenie odpowiedzi na leczenie. Eliminacja choroby resztkowej stanowi istotny czynnik rokowniczy długotrwałej odpowiedzi na immunochemioterapię. Ponadto znaczenie oceny MRD zwiększa się w związku z możliwością indywidualizacji leczenia CLL oraz oceny jego skuteczności bez konieczności długoletniej obserwacji chorych. W niniejszej pracy podsumowano znaczenie kliniczne oceny MRD w CLL ze szczególnym uwzględnieniem metodyki opartej na FLC. Słowa kluczowe: przewlekła białaczka limfocytowa, diagnostyka, minimalna choroba resztkowa, immunochemioterapia, cytometria przepływowa

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A single tube 10‐color flow cytometry assay optimizes detection of minimal residual disease in chronic lymphocytic leukemia

Cited by 26 publications

References 17 publications

Minimal residual disease monitoring in chronic lymphocytic leukaemia patients. A comparative analysis of flow cytometry and ASO IgH RQ‐PCR

Minimal residual disease monitoring in chronic lymphocytic leukaemia patients. A comparative analysis of flow cytometry and ASO IgH RQ‐PCR

Reference intervals for leukocyte subsets in adults: Results from a population‐based study using 10‐color flow cytometry

Znaczenie oceny minimalnej choroby resztkowej u chorych na przewlekłą białaczkę limfocytową

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