Rapid method for distinguishing clonal from polyclonal B cell populations in surgical biopsy specimens.

McCarthy, Keith; Sloane, J P; Wiedemann, Leanne M.

doi:10.1136/jcp.43.5.429

Cited by 172 publications

(62 citation statements)

References 14 publications

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“…On the other hand, the rapid and less labor-intensive PCR continues to gain popularity as a technique for the assessment of clonality in lymphoproliferative disorders. 1,2 The results obtained with antigen receptor gene PCR correlate well with the results of SBH. [3][4][5] PCR-based assays have additional advantages over SBH analysis.…”

Section: Polymerase Chain Reaction (Pcr)-based Analysis For Detectingsupporting

confidence: 69%

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

Elenitoba-Johnson¹,

Bohling²,

Mitchell³

et al. 2000

The Journal of Molecular Diagnostics

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Polymerase chain reaction (PCR)-based analysis for detecting immunoglobulin heavy chain gene (IgH)rearrangements in lymphoproliferative disorders is well established. The presence of one or two discrete bands is interpreted as a monoclonal proliferation, whereas a smear pattern represents a polyclonal population. Prompted by our observation of discrete bands in histologically reactive processes with a relative paucity of B cells, we sought to determine whether low numbers of B cells in biopsy specimens could artifactually produce pseudomonoclonal bands. We performed IgH PCR analysis on serially diluted DNA samples from 5 B cell non-Hodgkin's lymphomas (B-NHLs), 5 reactive lymph nodes, 5 reactive tonsils and 10 microdissected germinal centers from a lymph node with follicular hyperplasia. We also assessed multiple aliquots of DNA samples from small biopsy specimens of reactive lymphocytic processes from the stomach (5 cases). PCR products were evaluated using high resolution agarose or polyacrylamide gels, and DNA sequencing was performed on IgH PCR products from two reactive germinal centers, which yielded monoclonal bands of identical size. The utility of molecular techniques such as Southern blot hybridization (SBH) analysis and polymerase chain reaction (PCR) in the assessment of clonality in lymphoproliferative disorders is well established. Although SBH is a fairly sensitive and specific method, its utility as a routine diagnostic tool is hampered by its requirement for high quality DNA, its labor-intensive nature, and its consequent long turnaround time. On the other hand, the rapid and less labor-intensive PCR continues to gain popularity as a technique for the assessment of clonality in lymphoproliferative disorders. 1,2 The results obtained with antigen receptor gene PCR correlate well with the results of SBH. [3][4][5] PCR-based assays have additional advantages over SBH analysis. PCR requires smaller quantities of DNA, and high molecular weight DNA is not necessary. Therefore, PCR has been applied to small biopsies and fixed paraffin-embedded tissues, which are generally unsuitable for SBH analysis. 6 PCR is extremely sensitive and can detect 1 positive cell in a background of 10 5 negative cells. Immunoglobulin heavy chain (IgH) PCR is capable of detecting 1 monoclonal B cell in a background of 10 2 to 10 3 polyclonal B cells. 7 The extreme sensitivity of PCR also constitutes a potential source for pitfalls in the interpretation of PCR-based antigen receptor gene rearrangement studies. Clearly, contamination is a frequent concern. Additionally, we have observed discrete bands in samples obtained from small biopsy specimens in which histological and immunophenotypic evaluation revealed a reactive process. We believe that the discrete bands generated in this situation are related to the paucity of B

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Section: Polymerase Chain Reaction (Pcr)-based Analysis For Detectingsupporting

confidence: 69%

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

Elenitoba-Johnson¹,

Bohling²,

Mitchell³

et al. 2000

The Journal of Molecular Diagnostics

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show abstract

“…Sections of paraffin-embedded tissue were deparaffinized with xylene for 30 minutes, then dehydrated with 100% and 70% ethanol for 20 minutes each, followed by washing with dH 2 0 twice and digested with proteinase K at 42°C overnight. The digested DNA solution was then boiled for approximately 10 minutes to inactivate proteinase K. Immunoglobulin heavy-chain gene (IgH) rearrangements were evaluated by polymerase chain reaction (PCR) using VH region Framework 3 (Fr3A) consensus primer and a JH consensus primer as reported elsewhere (22). Primer sequences for IgH-PCR were JH (5'-ACC TGA GGA GAC GGT GAC C-3') and Fr3A (5'-ACA CGG CTA TGT ATT ACT GT-3').…”

Section: Molecular Studiesmentioning

confidence: 99%

An Immunophenotypic and Molecular Study of Primary Large B-Cell Lymphoma of Bone

et al. 2001

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“…A marker was positive when expressed by of IgH gene rearrangement (IgH-PCR). [6][7][8][9][10][11][12] Results from the two Ͼ20% of leukemic cells. techniques were similar.…”

Section: Immunophenotypingmentioning

confidence: 99%

Prognostic significance of immunoglobulin heavy chain gene rearrangement in patients with acute myelogenous leukemia

et al. 1997

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Rapid method for distinguishing clonal from polyclonal B cell populations in surgical biopsy specimens.

Abstract: The polymerase chain reaction (PCR)

Cited by 172 publications

References 14 publications

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

PCR Analysis of the Immunoglobulin Heavy Chain Gene in Polyclonal Processes Can Yield Pseudoclonal Bands as an Artifact of Low B Cell Number

An Immunophenotypic and Molecular Study of Primary Large B-Cell Lymphoma of Bone

Prognostic significance of immunoglobulin heavy chain gene rearrangement in patients with acute myelogenous leukemia

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