Bone marrow minimal disseminated disease (MDD) and minimal residual disease (MRD) in childhood T-cell lymphoblastic lymphoma stage III, detected by flow cytometry (FC) and real-time quantitative polymerase chain reaction (RQ-PCR)

2009 ◽  
Vol 52 (1) ◽  
pp. 20-25 ◽  
Author(s):  
Batia Stark ◽  
Smadar Avigad ◽  
Drorit Luria ◽  
Sigal Manor ◽  
Tali Reshef-Ronen ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Residual Disease ◽  
Quantitative Polymerase Chain Reaction ◽  
Lymphoblastic Lymphoma ◽  
Chain Reaction ◽  
Minimal Disseminated Disease ◽  
Polymerase Chain ◽  
Disseminated Disease ◽  
Minimal Residual

scholarly journals CURRENT STRATEGIES FOR THE DETECTION OF MINIMAL RESIDUAL DISEASE IN CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA

2016 ◽  
Vol 8 ◽  
pp. 2016024 ◽  
Author(s):  
Juliana Maria Camargos Rocha ◽  
Sandra Guerra Xavier ◽  
Marcelo Eduardo Lima Souza ◽  
Juliana Godoy Assumpção ◽  
Mitiko Murao ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
High Risk ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Minimal Residual

Acute lymphoblastic leukemia (ALL) is the most common cancer in children. Current treatment strategies for childhood ALL result in long term remission for approximately 90% of patients. However, therapeutic response is worse among those who relapse. Several risk stratification approaches based on clinical and biological aspects have been proposed in order to intensify treatment in patients with high risk of relapse and reduce toxicity on those with greater probability of cure.The detection of residual leukemic cells (minimal residual disease, MRD) is the most important prognostic factor to identify high risk patients, allowing redefinition of chemotherapy. In the last decades, several standardized research protocols evaluated MRD using immunophenotyping by flow cytometry and/or real time quantitative polymerase chain reaction at different time points during treatment. Both methods are highly sensitive (10-3 a 10-5), but expensive, complex, and, because of that, require qualified staff and frequently are restricted to reference centers.The aim of this article was to review technical aspects of immunophenotyping by flow cytometry and real time quantitative polymerase chain reaction to evaluate MRD in ALL. 

Clonotypic polymerase chain reaction confirms minimal residual disease in CLL nodular PR: results from a sequential treatment CLL protocol

Blood ◽  
2001 ◽  
Vol 97 (7) ◽  
pp. 1929-1936 ◽  
Author(s):  
Ariela Noy ◽  
Ravi Verma ◽  
Martha Glenn ◽  
Peter Maslak ◽  
Zia U. Rahman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
Lymphocytic Leukemia ◽  
High Dose ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Minimal Residual

Abstract Patient–tumor-specific oligonucleotides were generated for the detection of minimal residual disease (MRD) in a highly specific and sensitive clonotypic polymerase chain reaction (cPCR). The clone-specific region of highest diversity, CDR-III, was PCR amplified and sequenced. Nested CDR-III clonotypic primers were used in a semi-nested cPCR with a sensitivity of at least 1 in 105cells. Patients with protocol-eligible Rai intermediate or high-risk chronic lymphocytic leukemia (CLL) received induction with fludarabine 25 mg/m2 per day for 5 days every 4 weeks for 6 cycles, followed by consolidative high-dose cyclophosphamide (1.5, 2.25, or 3g/m2). cPCR was performed on peripheral blood and bone marrow mononuclear cells. All 5 patients achieving a clinical partial remission (PR) studied by cPCR were positive. Five patients achieved nodular PR (nPR) (residual nodules or suspicious lymphocytic infiltrates in a bone marrow biopsy as the sole suggestion of residual disease). Five of 5 patients with nPR were cPCR positive. In contrast, flow cytometry for CD5–CD19 dual staining and κ–λ clonal excess detected MRD in only 3 of the same 5 nPR patients, all of whom were cPCR positive, and immunohistochemistry detected MRD in only 1 of 4 assessable patients. Three of 7 CR patients evaluable by cPCR had MRD. Only 1 CR patient had MRD by flow cytometry; that patient was also cPCR positive. These data support the conclusions that nodular PR in CLL represents MRD and that clonotypic PCR detects MRD in CLL more frequently than flow cytometry or immunohistochemistry.

scholarly journals Mesenchymal Neuroblastoma Cells Are Undetected by Current mRNA Marker Panels: The Development of a Specific Neuroblastoma Mesenchymal Minimal Residual Disease Panel

2019 ◽  
pp. 1-11
Author(s):  
Esther M. van Wezel ◽  
Lieke M.J. van Zogchel ◽  
Jalenka van Wijk ◽  
Ilse Timmerman ◽  
Ngoc-Kim Vo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Peripheral Blood ◽  
Peripheral Blood Stem Cell ◽  
Residual Disease ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Mrna Markers ◽  
Minimal Residual

PURPOSE Patients with neuroblastoma in molecular remission remain at considerable risk for disease recurrence. Studies have found that neuroblastoma tissue contains adrenergic (ADRN) and mesenchymal (MES) cells; the latter express low levels of commonly used markers for minimal residual disease (MRD). We identified MES-specific MRD markers and studied the dynamics of these markers during treatment. PATIENTS AND METHODS Microarray data were used to identify genes differentially expressed between ADRN and MES cell lines. Candidate genes were then studied using real-time quantitative polymerase chain reaction in cell lines and control bone marrow and peripheral blood samples. After selecting a panel of markers, serial bone marrow, peripheral blood, and peripheral blood stem cell samples were obtained from patients with high-risk neuroblastoma and tested for marker expression; survival analyses were also performed. RESULTS PRRX1, POSTN, and FMO3 mRNAs were used as a panel for specifically detecting MES mRNA in patient samples. MES mRNA was detected only rarely in peripheral blood; moreover, the presence of MES mRNA in peripheral blood stem cell samples was associated with low event-free survival and overall survival. Of note, during treatment, serial bone marrow samples obtained from 29 patients revealed a difference in dynamics between MES mRNA markers and ADRN mRNA markers. Furthermore, MES mRNA was detected in a higher percentage of patients with recurrent disease than in those who remained disease free (53% v 32%, respectively; P = .03). CONCLUSION We propose that the markers POSTN and PRRX1, in combination with FMO3, be used for real-time quantitative polymerase chain reaction–based detection of MES neuroblastoma mRNA in patient samples because these markers have a unique pattern during treatment and are more prevalent in patients with poor outcome. Together with existing markers of MRD, these new markers should be investigated further in large prospective studies.

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