Comparative analysis of detection systems for evaluation of PCR amplified immunoglobulin heavy-chain gene rearrangements

Diagn Mol Pathol. 1997 Jun;6(3):140-6. doi: 10.1097/00019606-199706000-00003.

Abstract

Four different detection systems were compared for evaluation of polymerase chain reaction (PCR) amplified immunoglobulin heavy-chain gene rearrangements in acute lymphoblastic leukemia (ALL) and non-Hodgkin's lymphoma (NHL) of B-cell lineage. In 63.0% of the fragments detected by ethidium bromide stained agarose gel electrophoresis (Agarose-EtBr) the sensitivity was insufficient to separate the specific clonal population from the background of normal B cells. Using polyacrylamide gel electrophoresis (PAGE), PAGE combined with single-strand conformation polymorphism (PAGE-SSCP) and PhastGel-SSCP (Phast-SSCP) analysis with silver staining, the resolution was improved and the majority of the inconclusive amplicons were elucidated. However, Phast-SSCP displayed a slightly higher detection level compared to PAGE and PAGE-SSCP. According to our findings PAGE-SSCP and Phast-SSCP were superior to agarose-EtBr and PAGE in detecting new emerging clones and clonal evolution.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Neoplasm / isolation & purification
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Gene Rearrangement, B-Lymphocyte, Heavy Chain / genetics*
  • Humans
  • Immunoglobulin Variable Region / genetics*
  • Lymphoma, Non-Hodgkin / genetics*
  • Lymphoma, Non-Hodgkin / immunology
  • Nucleic Acid Amplification Techniques*
  • Polymerase Chain Reaction
  • Polymorphism, Single-Stranded Conformational
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / genetics*
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / immunology
  • Sensitivity and Specificity
  • Silver Staining

Substances

  • DNA, Neoplasm
  • Immunoglobulin Variable Region