Quantitative assessment of contaminating tumor cells in autologous peripheral blood stem cells of B-cell non-Hodgkin lymphomas using immunoglobulin heavy chain gene allele-specific oligonucleotide real-time quantitative-polymerase chain reaction

Leuk Res. 2003 Oct;27(10):925-34. doi: 10.1016/s0145-2126(03)00049-3.

Abstract

A real-time quantitative-polymerase chain reaction (RQ-PCR) targeting the immunoglobulin heavy chain (IgH) gene has been used for the quantification of minimal residual disease (MRD) in B-cell hematological malignancies. In non-Hodgkin lymphoma (NHL), experimental costs are increased, as a large number of primer-probe sets are required because of diversity, due to somatic and ongoing mutations of the IgH gene. We developed an allele-specific oligonucleotide (ASO) combined with a germline consensus probe-based RQ-PCR assay and examined MRD in peripheral blood stem cells (PBSC). The IgH consensus probes were adapted in seven (50%) of 14 amplifiable cases. Patients with heavily contaminating tumor cells in PBSC relapsed after PBSC transplantation. Our strategy will contribute to the development of a cost-efficient, precisely quantitative and systemic detection assay for MRD in NHL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Alleles
  • Base Sequence
  • Consensus Sequence
  • DNA Primers / economics
  • DNA Primers / standards
  • Female
  • Genes, Immunoglobulin / genetics*
  • Humans
  • Lymphoma, B-Cell / therapy*
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Neoplasm, Residual / diagnosis*
  • Neoplastic Cells, Circulating / pathology*
  • Oligonucleotides
  • Peripheral Blood Stem Cell Transplantation / methods*
  • Peripheral Blood Stem Cell Transplantation / standards
  • Polymerase Chain Reaction / economics
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Sensitivity and Specificity
  • Transplantation, Autologous

Substances

  • DNA Primers
  • Oligonucleotides