Prognostic Significance of Molecular Follow-up by Qualitative and Real-Time Quantitative Reverse-Transcriptase Polymerase Chain Reaction in TEL/AML1–Positive Childhood Acute Lymphoblastic Leukemia

2007 ◽  
Vol 1 (5) ◽  
pp. 298-303
Author(s):  
Gisella Volpe ◽  
Nicoletta Bertorello ◽  
Elena Barisone ◽  
Stefano Ulisciani ◽  
Enrico Gottardi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Lymphoblastic Leukemia ◽  
Prognostic Significance ◽  
Childhood Acute Lymphoblastic Leukemia ◽  
Chain Reaction ◽  
Polymerase Chain

Frequency of two transcripts of BCR-ABL 1 fusion gene (p210 and p190) in acute lymphoblastic leukemia and chronic myeloid leukemia by reverse transcriptase polymerase chain reaction

2021 ◽  
Vol 9 (1) ◽  
pp. 12-15
Author(s):  
Maria Altaf ◽  
Saleem Ahmed Khan ◽  
Ayesha Khan ◽  
Ammara Arsalan
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Fusion Gene ◽  
Lymphoblastic Leukemia ◽  
Prognostic Significance ◽  
Myelogenous Leukemia ◽  
Chain Reaction ◽  
Polymerase Chain

Objective: To determine the frequency of two transcripts of BCR-ABL 1 fusion gene (P210 & P190) in acute lymphoblastic leukemia and chronic myelogenous leukemia by reverse transcriptase polymerase chain reaction. Study design: A cross-sectional study. Place and duration of study: The Armed Forces Institute of Pathology (AFIP), Rawalpindi, from December 2012 to January 2014. Methodology: 147 diagnosed patients of CML and ALL were subjected to real time reverse transcriptase polymerase chain reaction. For PCR,2-3ml of venous blood in EDTA was collected .RNA extraction was done by Trizol Reagent LS (MRC, USA) and cDNA was synthesized using reverse transcriptase and gene specific primer. Real time- PCR was done on ABI-7500.The positive samples were identified when fluorescence exceeded threshold limit. Results: Out of 147 samples, 85 were with diagnosis of CML. All of them (100%) showed P210. 58 patients had diagnosis of ALL. Out of these BCR-ABL1 is detected in 11(18.9%) patients. 9(81.8%) expressed P190 whereas P210 was present in 2(18.1%) patients. 4 patients in lymphoid blast phase of CML were identified. 3(75%) of them had PP210 and 1(25%) shows both (P190 & P210) the transcripts. Conclusion: All CML patients expressed P210 transcript whereas ALL patients mostly showed P190. These transcripts expressing specific tyrosine kinase protein have diagnostic as well prognostic significance.

scholarly journals Long-term follow-up of residual disease in acute lymphoblastic leukemia patients in complete remission using clonogeneic IgH probes and the polymerase chain reaction

Blood ◽  
1993 ◽  
Vol 82 (5) ◽  
pp. 1618-1625 ◽  
Author(s):  
Y Nizet ◽  
S Van Daele ◽  
P Lewalle ◽  
JL Vaerman ◽  
M Philippe ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Complete Remission ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Clonal Evolution ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract We sequentially studied bone marrow (BM) samples of 25 patients in complete remission of an acute lymphoblastic leukemia (ALL) using a simplified polymerase chain reaction (PCR) strategy (direct use of the PCR product as a clonogenic probe recognizing rearranged Ig heavy chain sequences) as a first approach. BM aspirates were serially investigated after obtention of a complete response. When sensitivity was less than 1:10(4), the PCR fragment was sequenced and a specific oligonucleotide was synthetized and used as a probe (five cases). Cases in which minimal residual disease (MRD) became undetectable were cross- controlled using either TCR delta rearrangement or a specific translocation to circumvent the problem of false-negative results arising from clonal evolution. The median follow-up was 30 months (3 to 51 months). Within the first 3 months of complete remission, MRD was detectable in 22 of 23 investigated patients and remained so in 19 of 21 patients examined at 6 months, regardless of the long-term clinical outcome. In patients remaining in complete remission at 30 months or more, two patterns of MRD emerged during the follow-up. Either it continuously decreased to ultimately become undetectable (five patients) or remained detectable (five patients) with an increase after discontinuation of treatment in two. In the eight patients who relapsed, MRD persisted throughout the clinical course, and eventually increased 3 to 12 months before relapse was clinically detectable. In one case, clonal evolution of the VDJ heavy chain region was observed and recurrence of MRD shown by the use of TCR delta rearrangement as a control. We conclude that the use of this simplified methodology is a valuable tool for the follow-up of MRD in a majority of ALL patients, though in a few cases, sequencing needs to be performed to achieve a relevant sensitivity. The possibility of clonal evolution requires a cross-control of any sample becoming negative whatever the initial rearrangement used to generate a probe. In patients on therapy, sequential search for MRD seems to be a good tool for predicting the long-term outcome. In addition, patients remaining positive at the time treatment is discontinued or with a high tumor burden after a few months therapy may be at a higher risk of subsequent relapse, although a longer follow-up is needed to answer this question.

Serial Follow-Up and the Prognostic Significance of Reverse Transcriptase-Polymerase Chain Reaction—Staged Sentinel Lymph Nodes From Melanoma Patients

2004 ◽  
Vol 22 (19) ◽  
pp. 3989-3996 ◽  
Author(s):  
Udai S. Kammula ◽  
Ronald Ghossein ◽  
Satyajit Bhattacharya ◽  
Daniel G. Coit
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Prognostic Significance ◽  
Recurrence Risk ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Recurrence Rates ◽  
Single Marker ◽  
Polymerase Chain

Purpose Reverse transcriptase-polymerase chain reaction (RT-PCR) may provide an extremely sensitive method for detection of occult nodal disease. We evaluated the role of a single-marker RT-PCR assay for tyrosinase mRNA in the detection of melanoma sentinel lymph node (SLN) metastases and correlated the results with long-term clinical outcome. Patients and Methods One hundred twelve patients who underwent SLN biopsy for melanoma were prospectively analyzed. SLNs were bivalved, with half of each specimen evaluated by histologic methods and the other half evaluated by nested RT-PCR for tyrosinase. Results Fifteen patients (13%) had histologically positive SLNs, all of whom were also positive by RT-PCR (HISTO+/PCR+). Thirty-nine patients (35%) had SLNs that were negative by both histology and RT-PCR (HISTO−/PCR−). Fifty-eight patients (52%) were histologically negative but upstaged with a positive RT-PCR result (HISTO−/PCR+). Initially, at a median follow-up of 42 months, recurrence rates among the three cohorts were statistically different (HISTO+/PCR+, 53%; HISTO−/PCR+, 14%; and HISTO−/PCR−, 0%). However, at a longer median follow-up (67 months), recurrence rates for the HISTO−/PCR+ (24%) and HISTO−/PCR− (15%) groups were no longer statistically different (P = .25). The median time to relapse between the HISTO−/PCR+ and HISTO−/PCR− groups differed by 10 months (31 v 41 months, respectively). Conclusion With extended follow-up of patients with histologically negative SLNs, detection of submicroscopic disease by tyrosinase RT-PCR does not define a subgroup that is at higher recurrence risk when compared with patients with RT-PCR–negative SLNs. Future studies evaluating molecular staging will require approximately 5 years of median follow-up to accurately define outcome for patients with occult melanoma metastases.

scholarly journals Long-term follow-up of residual disease in acute lymphoblastic leukemia patients in complete remission using clonogeneic IgH probes and the polymerase chain reaction

Blood ◽  
1993 ◽  
Vol 82 (5) ◽  
pp. 1618-1625 ◽  
Author(s):  
Y Nizet ◽  
S Van Daele ◽  
P Lewalle ◽  
JL Vaerman ◽  
M Philippe ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Lymphoblastic Leukemia ◽  
Complete Remission ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Clonal Evolution ◽  
Chain Reaction ◽  
Polymerase Chain

We sequentially studied bone marrow (BM) samples of 25 patients in complete remission of an acute lymphoblastic leukemia (ALL) using a simplified polymerase chain reaction (PCR) strategy (direct use of the PCR product as a clonogenic probe recognizing rearranged Ig heavy chain sequences) as a first approach. BM aspirates were serially investigated after obtention of a complete response. When sensitivity was less than 1:10(4), the PCR fragment was sequenced and a specific oligonucleotide was synthetized and used as a probe (five cases). Cases in which minimal residual disease (MRD) became undetectable were cross- controlled using either TCR delta rearrangement or a specific translocation to circumvent the problem of false-negative results arising from clonal evolution. The median follow-up was 30 months (3 to 51 months). Within the first 3 months of complete remission, MRD was detectable in 22 of 23 investigated patients and remained so in 19 of 21 patients examined at 6 months, regardless of the long-term clinical outcome. In patients remaining in complete remission at 30 months or more, two patterns of MRD emerged during the follow-up. Either it continuously decreased to ultimately become undetectable (five patients) or remained detectable (five patients) with an increase after discontinuation of treatment in two. In the eight patients who relapsed, MRD persisted throughout the clinical course, and eventually increased 3 to 12 months before relapse was clinically detectable. In one case, clonal evolution of the VDJ heavy chain region was observed and recurrence of MRD shown by the use of TCR delta rearrangement as a control. We conclude that the use of this simplified methodology is a valuable tool for the follow-up of MRD in a majority of ALL patients, though in a few cases, sequencing needs to be performed to achieve a relevant sensitivity. The possibility of clonal evolution requires a cross-control of any sample becoming negative whatever the initial rearrangement used to generate a probe. In patients on therapy, sequential search for MRD seems to be a good tool for predicting the long-term outcome. In addition, patients remaining positive at the time treatment is discontinued or with a high tumor burden after a few months therapy may be at a higher risk of subsequent relapse, although a longer follow-up is needed to answer this question.

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