Author(s): Tarusawa M, Yashima A, Endo M, Maesawa C
We developed an assay using a real-time quantitative polymerase chain reaction (RQ-PCR) for the quantitative assessment of minimal residual disease (MRD) in childhood lymphoid malignancies by using a consensus V-region probe combining a allele-specific oligonucleotide (ASO) reverse primer. Our strategy employs a set consisting of a consensus V-region probe, an ASO reverse primer, and a patient-specific forward primer for clonal antigen-receptor (IgH, immunoglobulin heavy chain; TCR, T-cell receptor) gene rearrangements (IgH-ASO and TCR-ASO RQ-PCR assays). The limit of detection in both assays was 5 copies of the target/10(5) cell equivalents. We tested the assays in 17 childhood malignancies (14 cases of acute lymphoblastic leukemia and 3 of non-Hodgkin's lymphoma). High correlation coefficients of the standard curves (>0.980) and PCR efficiency (>0.95) were achieved with all primer/probe sets. In 2 (12%) of the 17 patients, ASO primers could not be designed because there was no junctional N-sequence. The quantitative data suggest that the copy number of clonal antigen receptors markedly decreased after induction therapy in 15 of 17 patients and that 1 patient relapsed and died of the disease. Consensus probes make it possible to examine a large number of patients with only a limited number of probes. The strategy used for IgH-ASO and TCR-ASO RQ-PCR assays is accurate and reliable in the clinical prospective study of MRD in childhood lymphoid malignancies.
Referred From: https://www.ncbi.nlm.nih.gov/pubmed/11939263
Author(s): LeBrun DP, Kamel OW, Cleary ML, Dorfman RF, Warnke RA
Author(s): Kodama T, Ohshima K, Nomura K, Taniwaki M, Nakamura N, et al.
Author(s): Yatabe Y, Nakamura S, Nakamura T, Seto M, Ogura M, et al.
Author(s): Hokama A, Tomoyose T, Yamamoto Y, Watanabe T, Hirata T, et al.
Author(s): Uchiyama M, Maesawa C, Yashima A, Itabashi T, Ishida Y, et al.
Author(s): Wan JH, Trainor KJ, Brisco MJ, Morley AA
Author(s): Ramasamy I, Brisco M, Morley A
Author(s): Aubin J, Davi F, Nguyen-Salomon F, Leboeuf D, Debert C, et al.
Author(s): Khaled S, Gotlieb V, Schuster IP, Saif MW
Author(s): Romaguera JE, Medeiros LJ, Hagemeister FB, Fayad LE, Rodriguez MA, et al.
Author(s): Medlicott SA, Brown HA, Roland B, Beck PL, Auer I, et al.
Author(s): Hamblin TJ, Davis Z, Gardiner A, Oscier DG, Stevenson FK