scholarly journals Prospective monitoring and quantitation of residual blasts in childhood acute lymphoblastic leukemia by polymerase chain reaction study of delta and gamma T-cell receptor genes

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1892-1902 ◽  
Author(s):  
H Cave ◽  
C Guidal ◽  
P Rohrlich ◽  
MH Delfau ◽  
A Broyart ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
T Cell Receptor ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Cell Receptor ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
B Lineage ◽  
Minimal Residual

We have developed a strategy based on polymerase chain reaction (PCR) for detecting all possible gamma T-cell receptor (gamma TCR) rearrangements and the most common delta TCR rearrangements found in B- lineage and T-acute lymphoblastic leukemia (T-ALL). The segments amplified from blasts are then directly sequenced to derive clonospecific probes. From a series of 45 patients aged 1 to 15 years (42 B-lineage ALL, 3 T-ALL), 35 (83%) could be followed for minimal residual disease with at least one clonospecific probe. Detection of clonal markers using clonospecific probes routinely allowed the detection of 1 to 10 blasts out of 10(5) cells as determined by serial dilutions of the initial samples. Residual disease was quantitated by a competitive PCR assay based on the coamplification of an internal standard. Twenty children were prospectively followed for periods varying from 7 to 30 months. In most children, a progressive decrease of the tumor load was observed, and blasts became undetectable within 6 months after the initiation of treatment. A slower kinetics of decrease in tumor cells was found in three children. These three patients relapsed with blasts that continued to display the initial clonospecific markers. Three other children had a central nervous system relapse despite the absence of detectable medullary residual disease. The use of both delta and gamma TCR genes as clonal markers, as well as simplification in the methods to detect and quantify residual blasts reported here, will allow the study of the large number of patients required to determine the role of the detection of minimal residual disease by PCR in the follow-up of childhood ALL.

scholarly journals Prospective monitoring and quantitation of residual blasts in childhood acute lymphoblastic leukemia by polymerase chain reaction study of delta and gamma T-cell receptor genes

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1892-1902 ◽  
Author(s):  
H Cave ◽  
C Guidal ◽  
P Rohrlich ◽  
MH Delfau ◽  
A Broyart ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
T Cell Receptor ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Cell Receptor ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
B Lineage ◽  
Minimal Residual

Abstract We have developed a strategy based on polymerase chain reaction (PCR) for detecting all possible gamma T-cell receptor (gamma TCR) rearrangements and the most common delta TCR rearrangements found in B- lineage and T-acute lymphoblastic leukemia (T-ALL). The segments amplified from blasts are then directly sequenced to derive clonospecific probes. From a series of 45 patients aged 1 to 15 years (42 B-lineage ALL, 3 T-ALL), 35 (83%) could be followed for minimal residual disease with at least one clonospecific probe. Detection of clonal markers using clonospecific probes routinely allowed the detection of 1 to 10 blasts out of 10(5) cells as determined by serial dilutions of the initial samples. Residual disease was quantitated by a competitive PCR assay based on the coamplification of an internal standard. Twenty children were prospectively followed for periods varying from 7 to 30 months. In most children, a progressive decrease of the tumor load was observed, and blasts became undetectable within 6 months after the initiation of treatment. A slower kinetics of decrease in tumor cells was found in three children. These three patients relapsed with blasts that continued to display the initial clonospecific markers. Three other children had a central nervous system relapse despite the absence of detectable medullary residual disease. The use of both delta and gamma TCR genes as clonal markers, as well as simplification in the methods to detect and quantify residual blasts reported here, will allow the study of the large number of patients required to determine the role of the detection of minimal residual disease by PCR in the follow-up of childhood ALL.

scholarly journals Detection of Philadelphia chromosome-positive acute lymphoblastic leukemia by polymerase chain reaction: possible eradication of minimal residual disease by marrow transplantation

Blood ◽  
1992 ◽  
Vol 79 (5) ◽  
pp. 1366-1370 ◽  
Author(s):  
K Miyamura ◽  
M Tanimoto ◽  
Y Morishima ◽  
K Horibe ◽  
K Yamamoto ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Minimal Residual Disease ◽  
Marrow Transplantation ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Minimal Residual

Abstract Minimal residual disease (MRD) in patients with Philadelphia chromosome- positive acute lymphoblastic leukemia (Ph1 ALL) who received allogeneic (n = 9) or autologous (n = 6) bone marrow transplantation (BMT) was evaluated by the polymerase chain reaction (PCR) for the bcr-abl transcript. Twelve patients received BMT at the time of hematologic and cytogenetic remission. However, MRD was detected in 8 of 10 patients evaluated. Seven patients, including three who had MRD before BMT, continue to have a disease-free survival 5 to 64 months after BMT. Twenty-one specimens obtained from these patients at various times after BMT did not show MRD. In three patients, MRD detected just before BMT seems to be eradicated by BMT protocol. The other eight patients developed cytogenetic or hematologic relapses 2 to 8 months after BMT. Seven of 14 samples from these patients demonstrated MRD, which preceded clinical relapse by 3 to 9 weeks. Thus, this technique for the detection of MRD appears to be useful for the more precise assessment of various antileukemia therapies and for early detection of leukemia recurrence.

Minimal Residual Disease in Childhood Acute Lymphoblastic Leukemia: A Comparative Study of Flow Cytometry and Quantitative Real-Time Polymerase Chain Reaction (RQ-PCR).

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4504-4504
Author(s):  
Ashish K. Misra ◽  
Mary Sartor ◽  
Rosemary Sutton ◽  
Nicola C. Venn ◽  
Shamira Cross ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Standard Medium ◽  
Chain Reaction ◽  
Medium Risk ◽  
Polymerase Chain ◽  
Minimal Residual

Abstract Approximately 25% of children with acute lymphoblastic leukemia (ALL) relapse after frontline therapy. Current stratification methods using clinical/biological criteria fail to identify a significant proportion of these children. Numerically more patients classified as standard/medium risk relapse than those with high-risk ALL. Early identification of this group with intensification of therapy may prevent this. We compare two techniques, multi-parameter flow cytometry (FCM) and real-time polymerase chain reaction (RQ-PCR) to monitor minimal residual disease (MRD) in bone marrow after initial therapy in newly diagnosed patients. We report our initial experience in patients with B-lineage ALL from a single institution (CHW) treated on the ANZ Haematology-Oncology Group Study VIII protocol for ALL. Multiparameter FCM was performed using two different four colour combinations of antibodies: CD19APC/CD45PerCP/CD10FITC/CD20PE and CD19APC/CD45PerCP/CD34FITC/CD9PE. A series of dual parameter displays were generated to define normal B cell differentiation pathways and this allowed the discrimination of malignant precursor B cells. RQ-PCR used clone specific primers and Taqman probes selected after sequencing the immunoglobulin heavy chain and/or T cell receptor gene rearrangements detected in the diagnosis DNA. The RQ-PCR assays had greater sensitivity and were highly reproducible. The timepoints used for analysis were Day 33 (D33) and D79, following induction and consolidation respectively. Of 45 patients, 44 (98%) had informative RQ-PCR markers: 12 (27%) with high MRD levels at D33 (>10−3) while 3 (6.7%) had high levels at D79. In contrast by FCM 5 (11.1%) had positive MRD (>0.1%) on D33 compared with 7 (15.6%) on D79. The 3 with high MRD on RQ-PCR at D79 were also identified by FCM. 2 of these 3 patients had been classified as standard/medium risk by conventional criteria. These two techniques may be complementary in identifying patients with significant MRD. Only a proportion of patients were identified by both methods. Long term follow up and ongoing recruitment will help delineate the optimal surveillance/stratification strategy by correlating these findings with outcome.

scholarly journals Prognostic significance of bi/oligoclonality in childhood acute lymphoblastic leukemia as determined by polymerase chain reaction

2001 ◽  
Vol 119 (5) ◽  
pp. 175-180 ◽  
Author(s):  
Carlos Alberto Scrideli ◽  
Ricardo Defavery ◽  
José Eduardo Bernardes ◽  
Luíz Gonzaga Tone
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Disease Free Survival ◽  
Chain Reaction ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Minimal Residual

CONTEXT: The CDR-3 region of heavy-chain immunoglobulin has been used as a clonal marker in the study of minimal residual disease in children with acute lymphoblastic leukemia. Southern blot and polymerase chain reaction studies have demonstrated the occurrence of bi/oligoclonality in a variable number of cases of B-lineage acute lymphoblastic leukemia, a fact that may strongly interfere with the detection of minimal residual disease. Oligoclonality has also been associated with a poorer prognosis and a higher chance of relapse. OBJECTIVES: To correlate bi/oligoclonality, detected by polymerase chain reaction in Brazilian children with B-lineage acute lymphoblastic leukemia with a chance of relapse, with immunophenotype, risk group, and disease-free survival. DESIGN: Prospective study of patients’ outcome. SETTING: Pediatric Oncology Unit of the University Hospital, Faculty of Medicine of Ribeirão Preto, University of São Paulo. PARTICIPANTS: 47 children with acute lymphoblastic leukemia DIAGNOSTIC TEST: Polymerase chain reaction using consensus primers for the CDR-3 region of heavy chain immunoglobulin (FR3A, LJH and VLJH) for the detection of clonality. RESULTS: Bi/oligoclonality was detected in 15 patients (31.9%). There was no significant difference between the groups with monoclonality and biclonality in terms of the occurrence of a relapse (28.1% versus 26.1%), presence of CALLA+ (81.2% versus 80%) or risk group (62.5% versus 60%). Disease-free survival was similar in both groups, with no significant difference (p: 0.7695). CONCLUSIONS: We conclude that bi/oligoclonality was not associated with the factors investigated in the present study and that its detection in 31.9% of the patients may be important for the study and monitoring of minimal residual disease.

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