Kappa deleting element as an alternative molecular target for minimal residual disease assessment by real-time quantitative PCR in patients with multiple myeloma

2012 ◽  
Vol 89 (4) ◽  
pp. 328-335 ◽  
Author(s):  
Noemí Puig ◽  
María E. Sarasquete ◽  
Miguel Alcoceba ◽  
Ana Balanzategui ◽  
María C. Chillón ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Residual Disease ◽  
Molecular Target ◽  
Disease Assessment ◽  
Real Time Quantitative Pcr ◽  
Minimal Residual

scholarly journals Application of Self-Quenched JH Consensus Primers for Real-Time Quantitative PCR of IGH Gene to Minimal Residual Disease Evaluation in Multiple Myeloma

2006 ◽  
Vol 8 (3) ◽  
pp. 364-370 ◽  
Author(s):  
Joaquin Martinez-Lopez ◽  
Pilar Martínez-Sanchez ◽  
Ramon Garcia-Sanz ◽  
Maria Eugenia Sarasquete ◽  
Rosa Ayala ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Residual Disease ◽  
Real Time Quantitative Pcr ◽  
Consensus Primers ◽  
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

PHOX2B Is a Novel and Specific Marker for Minimal Residual Disease Testing in Neuroblastoma

2008 ◽  
Vol 26 (33) ◽  
pp. 5443-5449 ◽  
Author(s):  
Janine Stutterheim ◽  
Annemieke Gerritsen ◽  
Lily Zappeij-Kannegieter ◽  
Ilona Kleijn ◽  
Rob Dee ◽  
...  
Keyword(s):  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Peripheral Blood ◽  
Residual Disease ◽  
Specific Marker ◽  
Real Time Quantitative Pcr ◽  
Normal Tissues ◽  
Specificity And Sensitivity ◽  
Minimal Residual

Purpose Polymerase chain reaction (PCR)–based detection of minimal residual disease (MRD) in neuroblastoma can be used to monitor therapy response and to evaluate stem cell harvests. Commonly used PCR markers, tyrosine hydroxylase (TH) and GD2 synthase, have expression in normal tissues, thus limiting MRD detection. To identify a more specific MRD marker, we tested PHOX2B. Patients and Methods To determine PHOX2B, TH, and GD2 synthase expression in normal tissues, it was measured by real-time quantitative PCR in samples of normal bone marrow (BM; n = 51), peripheral blood (PB; n = 37), and peripheral-blood stem cells (PBSCs; n = 24). Then, 289 samples of 101 Dutch patients and 47 samples of 43 German patients were tested for PHOX2B and TH; these samples included 52 tumor, 214 BM, 32 BM, and 38 PBSC harvests. Of the 214 BM samples, 167 were compared with cytology, and 47 BM samples were compared with immunocytology (IC). Results In contrast to TH and GD2 synthase, PHOX2B was not expressed in any of the normal samples. In patient samples, PHOX2B was detected in 32% cytology-negative and in 14% IC-negative samples and in 94% of cytology-positive and in 90% of IC-positive BM samples. Overall, PHOX2B was positive in 43% compared with 31% for TH. In 24% of all samples, TH expression was inconclusive, which is similar to expression found in normal tissues. In 42% of these samples, PHOX2B expression was positive. Conclusion PHOX2B is superior to TH and GD2 synthase in specificity and sensitivity for MRD detection of neuroblastoma by using real-time quantitative PCR. We propose to include PHOX2B in additional prospective MRD studies in neuroblastoma alongside TH and other MRD markers.

scholarly journals Detecting Minimal Residual Disease in Neuroblastoma: The Superiority of a Panel of Real-Time Quantitative PCR Markers

2009 ◽  
Vol 55 (7) ◽  
pp. 1316-1326 ◽  
Author(s):  
Janine Stutterheim ◽  
Annemieke Gerritsen ◽  
Lily Zappeij-Kannegieter ◽  
Bilgehan Yalcin ◽  
Rob Dee ◽  
...  
Keyword(s):  
Real Time ◽  
Minimal Residual Disease ◽  
Quantitative Pcr ◽  
Residual Disease ◽  
Therapy Response ◽  
Pcr Markers ◽  
Real Time Quantitative Pcr ◽  
Monitoring Therapy ◽  
Additional Value ◽  
Minimal Residual

Abstract Background: PCR-based detection of minimal residual disease (MRD) in neuroblastoma (NB) patients can be used for initial staging and monitoring therapy response in bone marrow (BM) and peripheral blood (PB). PHOX2B has been identified as a sensitive and specific MRD marker; however, its expression varies between tumors. Therefore, a panel of markers could increase sensitivity. Methods: To identify additional MRD markers for NB, we selected genes by comparing SAGE (serial analysis of gene expression) libraries of healthy and NB tissues followed by extensive real-time quantitative PCR (RQ-PCR) testing in samples of tumors (n = 56), control BM (n = 51), PB (n = 37), and cell subsets. The additional value of a panel was determined in 222 NB samples from 82 Dutch stage 4 NB patients (54 diagnosis BM samples, 143 BM samples during/after treatment, and 25 PB samples). Results: We identified 2 panels of specific RQ-PCR markers for MRD detection in NB patients: 1 for analysis of BM samples (PHOX2B, TH, DDC, CHRNA3, and GAP43) and 1 for analysis of PB samples (PHOX2B, TH, DDC, DBH, and CHRNA3). These markers all showed high expression in NB tumors and no or low expression in control BM or PB samples. In patients’ samples, the PHOX2B marker detected most positive samples. In PB samples, however, 3 of 7 PHOX2B-negative samples were positive for 1 or more markers, and in BM examinations during treatment, 7% (6 of 86) of the PHOX2B-negative samples were positive for another marker. Conclusions: Because of differences in the sensitivities of the markers in BM and PB, we advise the use of 2 different panels to detect MRD in these compartments.

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