Comparison of capillary electrophoresis and polyacrylamide gel electrophoresis for the evaluation of T and B cell clonality by polymerase chain reaction.
Polymerase chain reaction (PCR) technique is widely used in the diagnosis of lymphoma, and PCR amplification products are typically detected by polyacrylamide gel electrophoresis (PAGE). However, the identification of small clonal populations, or the distinction of clonal PCR products in a polyclonal milieu remains difficult, requiring technically demanding alterations to gel analysis. This study describes an alternative approach using a capillary electrophoresis (CE) system to produce an accurately sized electropherogram. A variety of patient samples were examined, including solid tissue, peripheral blood, bone marrow aspirates, and paraffin-embedded tissue. A total of 28 samples were evaluated by PCR for B-cell clonality by detection of immunoglobulin heavy chain gene rearrangement and 29 samples for T-cell clonality by detection of T-cell gamma locus gene rearrangement. Standard 10% PAGE analysis of PCR products was compared with CE. There was a 100% concordance in the assessment of both B-cell and T-cell clonality. Dilution studies with the SUP-B15 cell line showed a detection limit of 0.03% for B-cell clonality and 0.05% for T-cell clonality using CE, versus 0.2% to 1%, respectively for PAGE. Automated, fluorescent analysis of PCR products by CE seems to be at least equally as effective as gel-based analysis for the detection of clonal B-cell and T-cell populations. Moreover. CE offers superior resolution and improved sensitivity, thus representing a significant improvement over traditional gel electrophoretic techniques in these regards.
Beaubier
N T
NT
Department of Pathology and Center for Molecular and Cellular Diagnostics, University of New Mexico Health Sciences Center, Albuquerque 87131, USA.
Hart
A P
AP
Bartolo
C
C
Willman
C L
CL
Viswanatha
D S
DS
D001402:B-Lymphocytes / Q000737:chemistry / Q000166:cytology*; D001773:Blood Cells / Q000737:chemistry / Q000166:cytology; D001854:Bone Marrow Cells / Q000737:chemistry / Q000166:cytology; D002999:Clone Cells / Q000737:chemistry / Q000166:cytology*; D004247:DNA / Q000235:genetics; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D005470:Fluorometry; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D019337:Hematologic Neoplasms / Q000097:blood / Q000473:pathology; D006801:Humans; D016246:Lymphocytes, Tumor-Infiltrating / Q000473:pathology; D009369:Neoplasms / Q000097:blood / Q000276:immunology / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000473:pathology; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012984:Software; D013601:T-Lymphocytes / Q000737:chemistry / Q000166:cytology*
D004247:DNA