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 Citations (Scopus)

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 publicationMethods in Molecular Biology
Pages151-167
Number of pages17
Volume999
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume999
ISSN (Print)10643745

Fingerprint

Immunoglobulin Heavy Chain Genes
Gene Rearrangement
Capillary Electrophoresis
B-Lymphocytes
Gels
Polymerase Chain Reaction
Immunoglobulin Heavy Chains
Immunophenotyping
Lymphoproliferative Disorders
Multiplex Polymerase Chain Reaction
Fluorescence In Situ Hybridization
Cytogenetics
Neoplasms
Immunohistochemistry
Technology

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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 Methods in Molecular Biology (Vol. 999, pp. 151-167). (Methods in Molecular Biology; Vol. 999). https://doi.org/10.1007/978-1-62703-357-2-10

Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis. / Fan, Hongxin; Robetorye, Ryan S.

Methods in Molecular Biology. Vol. 999 2013. p. 151-167 (Methods in Molecular Biology; Vol. 999).

Research output: Chapter in Book/Report/Conference proceedingChapter

Fan, H & Robetorye, RS 2013, Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis. in Methods in Molecular Biology. vol. 999, Methods in Molecular Biology, vol. 999, pp. 151-167. https://doi.org/10.1007/978-1-62703-357-2-10
Fan, Hongxin ; Robetorye, Ryan S. / Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis. Methods in Molecular Biology. Vol. 999 2013. pp. 151-167 (Methods in Molecular Biology).
@inbook{dca73c445bc14266be6fc2db28160d58,
title = "Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis",
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.",
keywords = "Capillary gel electrophoresis, Clonality testing, Immunoglobulin heavy chain gene, PCR",
author = "Hongxin Fan and Robetorye, {Ryan S.}",
year = "2013",
doi = "10.1007/978-1-62703-357-2-10",
language = "English (US)",
isbn = "9781627033565",
volume = "999",
series = "Methods in Molecular Biology",
pages = "151--167",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Detection of clonal immunoglobulin heavy chain gene rearrangements by the polymerase chain reaction and capillary gel electrophoresis

AU - Fan, Hongxin

AU - Robetorye, Ryan S.

PY - 2013

Y1 - 2013

N2 - 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.

AB - 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.

KW - Capillary gel electrophoresis

KW - Clonality testing

KW - Immunoglobulin heavy chain gene

KW - PCR

UR - http://www.scopus.com/inward/record.url?scp=84883248171&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84883248171&partnerID=8YFLogxK

U2 - 10.1007/978-1-62703-357-2-10

DO - 10.1007/978-1-62703-357-2-10

M3 - Chapter

SN - 9781627033565

VL - 999

T3 - Methods in Molecular Biology

SP - 151

EP - 167

BT - Methods in Molecular Biology

ER -