Single-cell PCR analysis of the immunoglobulin heavy-chain CDR3 region for the diagnosis of leptomeningeal involvement of B-cell malignancies using standard cerebrospinal fluid cytospins.
The diagnosis of leptomeningeal B-cell malignancies is based on the identification of malignant B cells in the cerebrospinal fluid (CSF). We have established a polymerase chain reaction (PCR) approach to characterize the clonally diverse gene encoding the immunoglobulin heavy-chain (IgH) third complementarity determining region (CDR3) of single B cells. We demonstrate that single-cell PCR is readily applicable to individual cells derived from routine CSF cytospins and is a powerful method to discriminate monoclonal neoplastic from polyclonal reactive B-cell responses. Single-cell PCR analysis, as a new tool for the diagnosis and monitoring of neoplastic meningitis associated with B-cell malignancies, is particularly important if cytology, immunocytochemistry, flow cytometry and automated gene scanning of CSF samples are unable to detect malignant monoclonal proliferation.
Hug
Andreas
A
Department of Neurology, University of Heidelberg, Im Neuenheimer Feld 400, D-69120 Heidelberg, Germany.
Storch-Hagenlocher
Brigitte
B
Haas
Juergen
J
Vogt-Schaden
Maria-Elisabeth
ME
Goldschmidt
Hartmut
H
Wildemann
Brigitte
B
D001402:B-Lymphocytes; D001483:Base Sequence; D002555:Cerebrospinal Fluid / Q000166:cytology; D022801:Complementarity Determining Regions / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D004273:DNA, Neoplasm / Q000032:analysis; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D016393:Lymphoma, B-Cell / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000235:genetics*; D008577:Meningeal Neoplasms / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
D022801:Complementarity Determining Regions; D004273:DNA, Neoplasm; D007143:Immunoglobulin Heavy Chains