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      Deep-sequencing approach for minimal residual disease detection in acute lymphoblastic leukemia.

      Blood
      Base Sequence, Child, Clonal Evolution, genetics, physiology, Genes, Immunoglobulin Heavy Chain, High-Throughput Nucleotide Sequencing, methods, Humans, Models, Biological, Molecular Diagnostic Techniques, Molecular Sequence Data, Neoplasm, Residual, Polymerase Chain Reaction, Precursor Cell Lymphoblastic Leukemia-Lymphoma, diagnosis, pathology, Prognosis, Sensitivity and Specificity

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          Abstract

          The persistence of minimal residual disease (MRD) during therapy is the strongest adverse prognostic factor in acute lymphoblastic leukemia (ALL). We developed a high-throughput sequencing method that universally amplifies antigen-receptor gene segments and identifies all clonal gene rearrangements (ie, leukemia-specific sequences) at diagnosis, allowing monitoring of disease progression and clonal evolution during therapy. In the present study, the assay specifically detected 1 leukemic cell among greater than 1 million leukocytes in spike-in experiments. We compared this method with the gold-standard MRD assays multiparameter flow cytometry and allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) using diagnostic and follow-up samples from 106 patients with ALL. Sequencing detected MRD in all 28 samples shown to be positive by flow cytometry and in 35 of the 36 shown to be positive by ASO-PCR and revealed MRD in 10 and 3 additional samples that were negative by flow cytometry and ASO-PCR, respectively. We conclude that this new method allows monitoring of treatment response in ALL and other lymphoid malignancies with great sensitivity and precision. The www.clinicaltrials.gov identifier number for the Total XV study is NCT00137111.

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