Droplet digital PCR for BCR/ABL(P210) detection of chronic myeloid leukemia: A high sensitive method of the minimal residual disease and disease progression

2018 ◽  
Vol 101 (3) ◽  
pp. 291-296 ◽  
Author(s):  
Wen-Jun Wang ◽  
Chao-Feng Zheng ◽  
Zhuang Liu ◽  
Yan-Hong Tan ◽  
Xiu-Hua Chen ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Disease Progression ◽  
Minimal Residual Disease ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Minimal Residual ◽  
Sensitive Method

scholarly journals Treatment-free remission in Chronic Myeloid Leukemia harboring atypical BCR-ABL1 transcripts.

2020 ◽  
Vol 12 (1) ◽  
pp. e2020066 ◽  
Author(s):  
Matteo Dragani ◽  
Jessica Petiti ◽  
Giovanna Rege-Cambrin ◽  
Enrico Gottardi ◽  
Filomena Daraio ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Tyrosine Kinase ◽  
Minimal Residual Disease ◽  
Myeloid Leukemia ◽  
Kinase Inhibitors ◽  
Residual Disease ◽  
Digital Pcr ◽  
Treatment Free Remission ◽  
Minimal Residual

Discontinuation of tyrosine kinase inhibitors (TKI) is the main goal today in the field of Philadelphia positive chronic myeloid leukemia (Ph + CML) and the criteria to attempt the interruption of therapy are well defined and rely on the possibility to regularly monitor the BCR-ABL1 transcript. Patients harboring atypical transcripts are automatically excluded from protocols due to the absence of a standardized method of quantification of their minimal residual disease (MRD). We report here the outcome of 6 patients with atypical transcripts with a long follow up whose MRD was followed in three cases with digital PCR during their treatment free remission (TFR).

scholarly journals Novel Multiplex Droplet Digital PCR Assays to Monitor Minimal Residual Disease in Chronic Myeloid Leukemia Patients Showing Atypical BCR-ABL1 Transcripts

2020 ◽  
Vol 9 (5) ◽  
pp. 1457 ◽  
Author(s):  
Jessica Petiti ◽  
Marco Lo Iacono ◽  
Matteo Dragani ◽  
Lucrezia Pironi ◽  
Cristina Fantino ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Nested Pcr ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Molecular Response ◽  
Highly Sensitive ◽  
Treatment Free Remission ◽  
Minimal Residual

BCR-ABL1 fusion transcript is the minimal residual disease marker in chronic myeloid leukemia; 2% of patients show unusual breakpoints generating atypical transcripts, not quantifiable by standardized real-time PCR (RT–PCR). Response monitoring is performed by non-quantitative NESTED PCR, useless for evaluating patients’ molecular remission, excluding them from treatment-free-remission protocols. Droplet digital PCR (ddPCR) is highly sensitive technology, allowing an absolute quantification independent of standard curves. Based on this, we have developed assays able to evaluate the molecular response in atypical patients. We designed new ddPCR-based molecular assays able to quantify atypical BCR-ABL1 transcripts, with a detection limit of 0.001%, validated in a cohort of 65 RNA from 11 patients. Fifty samples were identified congruently by ddPCR and NESTED PCR (40 positives and 10 negatives for atypical BCR–ABL1 transcript), while 11 positive samples were detected only by ddPCR. Our results highlight ddPCR usefulness, primarily when the BCR–ABL1/ABL1 level is less than 1.5% and NESTED PCR results are often inaccurate. Furthermore, we identified 3 patients who maintained a deep molecular response for at least one year, who could be considered good candidates for treatment-free remission approaches. Here, we describe a new promising molecular approach, highly sensitive, to monitor atypical BCR–ABL1 patients, paving the foundation to include them in treatment-free remission protocols.

Endogenous Erythroid Colony Formation in Chronic Myeloid Leukemia: A Recurrent Finding Associated with Persistent Minimal Residual Disease Under Imatinib

2013 ◽  
Vol 22 (23) ◽  
pp. 3043-3051 ◽  
Author(s):  
Elisa Einwallner ◽  
Eva Jaeger ◽  
Gerlinde Mitterbauer-Hohendanner ◽  
Martin Bilban ◽  
Ingrid Simonitsch-Klupp ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Minimal Residual Disease ◽  
Colony Formation ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Minimal Residual

scholarly journals Manifold-assisted Reverse Transcription-PCR with Real-Time Detection for Measurement of the BCR-ABL Fusion Transcript in Chronic Myeloid Leukemia Patients

2000 ◽  
Vol 46 (7) ◽  
pp. 913-920 ◽  
Author(s):  
Gisela Barbany ◽  
Anette Hagberg ◽  
Ulla Olsson-Strömberg ◽  
Bengt Simonsson ◽  
Ann-Christine Syvänen ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Real Time ◽  
Minimal Residual Disease ◽  
Reverse Transcription ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Detection System ◽  
Fusion Transcript ◽  
Mrna Quantification ◽  
Minimal Residual

Abstract Background: BCR-ABL fusion mRNA expression in bone marrow or peripheral blood can be used as a measure of minimal residual disease in patients with chronic myeloid leukemia (CML). Methods: We used an oligo(dT)-coated manifold support to capture the mRNA directly from the cell lysate. After reverse transcription, the cDNA was eluted from the manifold support, and BCR-ABL and GAPDH mRNAs were quantified in real time using the TaqMan fluorogenic detection system. Results: The detection limit of the method was one positive K562 cell among 105 negative cells. GAPDH was chosen as a reference gene based on the low variation between samples from different stages of the disease and the low signal in the absence of reverse transcription. The day-to-day variation of the method (CV) was 32%. In 43 blood samples from 13 CML patients, mRNA quantification agreed well with cytogenetic data. Conclusions: The proposed procedure constitutes a reproducible and sensitive BCR-ABL mRNA quantification method and is suitable to monitor minimal residual disease in CML patients.

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