A competitive PCR-based method using TCRD, TCRG and IGH rearrangements for rapid detection of patients with high levels of minimal residual disease in acute lymphoblastic leukemia

Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED-2 Concerted Action BMH4-CT98-3936

“…[201][202][203] In lymphoid malignancies, sensitive and quantitative detection of MRD can be achieved by real-time quantitative PCR analysis of patient-specific junctional regions of rearranged Ig/TCR genes. 68,113,195,204 This requires rapid and easy detection and identification of one (or two) Ig/TCR gene rearrangements in each patient with a lymphoid malignancy. It will be clear from the preceding sections that the BIOMED-2 multiplex PCR tubes are perfectly suited for this purpose.…”

Standardized MRD quantification in European ALL trials: Proceedings of the Second International Symposium on MRD assessment in Kiel, Germany, 18–20 September 2008

“…An early simplified approach with limited sensitivity (B10 À3 ) based on competitive PCR and Genescan analysis allowed the detection of patients with high levels of residual blasts. 45 The implementation of real-time quantitative (RQ)-PCR techniques was a major advance, allowing for quantification during the early exponential phase of PCR amplification and eliminating variations during later postexponential phases of the PCR reaction and during post-PCR manipulation of samples. 28,39,40,46,47 Right from the start, standardization of RQ-PCR analysis and data interpretation was a major aim of European collaborations.…”

Clonality profile in relapsed precursor-B-ALL children by GeneScan and sequencing analyses. Consequences on minimal residual disease monitoring

“…Scanning results of fluorescence-dye-labelled PCR products by GSA were interpreted as follows: one or two striking peaks indicated a dominant leukaemic clone, while polyclonality was characterized by a number of peaks arranged in a normal distribution. 20,[26][27][28] A sensitivity test was also performed, diluting diagnostic or relapse samples in MNC DNA: clonal rearrangements of a characteristic size identified at presentation and relapse were detectable at a dilution of 1 Â 10 À3 in more than 50% of cases, and at a level of 5 Â 10 À3 in 89% of samples.…”

“…16,17 The high-resolution PCR fragment analysis using GeneScan technique (PCR-GSA) is an accurate procedure in determining the size of PCR products and in discriminating between polyclonal and clonal DNA banding profiles. [18][19][20] In this study, we carried out complete GeneScan and sequence analyses of every Ig and TCR gene rearrangement independently found at diagnosis and relapse in a homogeneous series of 53 childhood precursor-B-ALL patients, in order to get detailed information on the evolution and the stability of potentially useful MRD-PCR targets.…”