Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis

Hongxin Fan, Ryan S. Robetorye

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

Although well-established diagnostic criteria exist for mature B-cell neoplasms, a definitive diagnosis of a B-cell lymphoproliferative disorder cannot always be obtained using more conventional techniques such as flow cytometric immunophenotyping, conventional cytogenetics, fluorescence in situ hybridization, or immunohistochemistry. However, because B-cell malignancies contain identically rearranged immunoglobulin heavy chain genes, the polymerase chain reaction (PCR) can be a fast, convenient, and dependable option to identify clonal B-cell processes. This chapter describes the use of PCR and capillary electrophoresis to identify clonal immunoglobulin heavy chain (IGH) variable and joining region (VH-JH) gene rearrangements (IGH VH-JH PCR) using a commercially available method employing multiple multiplex PCR tubes that was originally developed as the result of a large European BIOMED-2 collaborative study (Invivoscribe Technologies). The core protocol involves the use of three separate master mix tubes that target the conserved framework (FR1, FR2, and FR3) and joining (J) regions of the IGH gene. Analysis of these three framework regions can detect approximately 88% of clonal IGH gene rearrangements.

Original languageEnglish (US)
Title of host publicationHematological Malignancies
Pages151-167
Number of pages17
DOIs
StatePublished - Sep 4 2013

Publication series

NameMethods in Molecular Biology
Volume999
ISSN (Print)1064-3745

Keywords

  • Capillary gel electrophoresis
  • Clonality testing
  • Immunoglobulin heavy chain gene
  • PCR

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis'. Together they form a unique fingerprint.

  • Cite this

    Fan, H., & Robetorye, R. S. (2013). Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis. In Hematological Malignancies (pp. 151-167). (Methods in Molecular Biology; Vol. 999). https://doi.org/10.1007/978-1-62703-357-2-10