scholarly journals A new highly sensitive real-time quantitative-PCR method for detection of BCR-ABL1 to monitor minimal residual disease in chronic myeloid leukemia after discontinuation of imatinib

PLoS ONE ◽  
2019 ◽  
Vol 14 (3) ◽  
pp. e0207170 ◽  
Author(s):  
Hiroaki Kitamura ◽  
Yoko Tabe ◽  
Tomohiko Ai ◽  
Koji Tsuchiya ◽  
Maiko Yuri ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Real Time Quantitative Pcr ◽  
Highly Sensitive ◽  
Pcr Method ◽  
Minimal Residual

scholarly journals A new highly sensitive real-time quantitative-PCR method for detection of BCR-ABL1 to monitor minimal residual disease in chronic myeloid leukemia after discontinuation of imatinib

2018 ◽  
Author(s):  
Hiroaki Kitamura ◽  
Yoko Tabe ◽  
Koji Tsuchiya ◽  
Maiko Yuri ◽  
Tomohiko Ai ◽  
...  
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Real Time ◽  
Minimal Residual Disease ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Quantitative Polymerase Chain Reaction ◽  
Philadelphia Chromosome ◽  
Molecular Response ◽  
Pcr Method ◽  
Minimal Residual

AbstractTyrosine kinase inhibitors (TKIs) targeting the BCR-ABL1 fusion protein, encoded by the Philadelphia chromosome, have drastically improved the outcomes for patients with chronic myeloid leukemia (CML). Although several real-time quantitative polymerase chain reaction (RQ-PCR) kits for the detection of BCR-ABL1 transcripts are commercially available, their accuracy and efficiency in laboratory practice require reevaluation. We have developed a new in-house RQ-PCR method to detect minimal residual disease (MRD) in CML cases. MRD was analyzed in 102 patients with CML from the DOMEST study, a clinical trial to study the rationale for imatinib mesylate discontinuation in Japan. The BCR-ABL1/ABL1 ratio was evaluated using the international standard (IS) ratio, where IS < 0.01% was defined as a major molecular response. At enrollment, BCR-ABL1 transcripts were undetectable in all samples using a widely-applied RQ-PCR method performed in the commercial laboratory, BML (BML Inc., Tokyo, Japan); however, the in-house method detected the BCR-ABL1 transcripts in five samples (5%) (mean IS ratio: 0.0062 ± 0.0010%). After discontinuation of imatinib, BCR-ABL1 transcripts were detected using the in-house RQ-PCR in 21 patients (21%) that were not positive using the BML method. Nineteen samples were also tested using a commercially available RQ-PCR assay kit with a detection limit of IS ratio, 0.0007% (ODK-1201, Otsuka Pharmaceutical Co., Tokyo, Japan). This method detected low levels of BCR-ABL1 transcripts in 14 samples (74%), but scored negative for five samples (26%) that were positive using the in-house method. These data suggest that our new in-house RQ-PCR method is effective for monitoring MRD in CML.

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.

scholarly journals Improved assessment of minimal residual disease in B cell malignancies using fluorogenic consensus probes for real-time quantitative PCR

Leukemia ◽  
2000 ◽  
Vol 14 (8) ◽  
pp. 1419-1425 ◽  
Author(s):  
M Brüggemann ◽  
J Droese ◽  
I Bolz ◽  
P Lüth ◽  
C Pott ◽  
...  
Keyword(s):  
B Cell ◽  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Residual Disease ◽  
B Cell Malignancies ◽  
Real Time Quantitative Pcr ◽  
Minimal Residual

Prognostic Value of Minimal Residual Disease by Real-Time Quantitative PCR in AML with CBFB-MYH11 Rearrangement: The French Experience.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3496-3496
Author(s):  
Romain Guièze ◽  
Aline Renneville ◽  
Jean-Michel Cayuela ◽  
Raouf Ben Abdelali ◽  
Nicolas Boissel ◽  
...  
Keyword(s):  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Residual Disease ◽  
Transcript Level ◽  
Prognostic Impact ◽  
Consolidation Therapy ◽  
Consolidation Treatment ◽  
Real Time Quantitative Pcr ◽  
Minimal Residual

Abstract Background. Despite the favorable prognosis of AML patients with inv(16)/t(16;16) leading to CBFB-MYH11 rearrangement, relapses still occur in about 40% of the cases. With the possible exception of receptor tyrosine kinase mutations, no pretreatment-factor can strongly predict risk of relapse. Methods. To explore minimal residual disease (MRD) prognostic impact, we prospectively monitored CBFB-MYH11 rearrangement by real-time quantitative PCR (RQ-PCR) in patients with inv(16)/t(16;16) treated in French cooperative trials; most patients were treated in Acute Leukemia French Association (ALFA) trials and Leucémies Aiguës Myéloblastiques de l’Enfant (LAME) Cooperative Groups trials. RQ-PCR was performed in three molecular French labs (CHU of Lille, Necker Hospital and St-Louis Hospital) according to the EAC procedure and results are expressed as CBFB-MYH11/100 ABL copies(%). Results. 61 AML patients with inv(16)/t(16;16) who had reached CR with one of the anthracyclin-aracytin regimens were analyzed. Median age was 37.5 years (range 2.1–76.5) and median baseline WBC 46 G/L (range 1.8–246). With a median follow-up of 23.3 months, median overall DFS was 23 months and median OS not reached (93.5% of patients alive). No initial clinical or hematological characteristic including age, gender and initial WBC was predictive of relapse in adults, but children had poorer DFS than adults (17.6 months vs not reached, p=0.03). Regarding chromosomal abnormalities in addition to inv(16)/t(16;16), trisomy 22 was the most frequent but no significant prognostic impact was observed. Of the 61 patients, MRD monitoring could be performed sequentially in 45. Pretreatment CBFB-MYH11 transcript level had no impact on DFS. However, after induction therapy, transcript levels >0.5% were significant predictors of poorer DFS (median 16.4 months vs not reached if <0.5%, p=0.002). Likewise, after first consolidation therapy course, transcript level >0.001% was also predictive of poorer DFS (median 22 months when >0.001% vs not reached when <0.001%; p=0.03). 34 paired analyses of bone marrow (BM) and peripheral blood (PB) samples revealed only moderate correlation (R2=0.86); especially for low MRD levels, CBFB-MYH11 transcript expression being generally higher (up to 78-fold) in BM samples than in PB samples. Conclusion. MRD monitoring by RQ-PCR appears to be a major prognostic factor of DFS in AML with CBFB-MYH11 rearrangement and could be a powerful tool to early identify good and bad responders which could have an impact on therapeutic decisions for consolidation therapy. DFS according to MRD level after first consolidation treatment DFS according to MRD level after first consolidation treatment

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