Polymerase chain reaction on cerebrospinal fluid cells in the detection of leptomeningeal involvement by B-cell lymphoma and leukaemia: a novel strategy and its implications.
In the search of B-cell lymphoma/leukaemia dissemination to cerebrospinal fluid (CSF), we used the highly sensitive semi-nested PCR (snPCR) for the analysis of IgH gene rearrangements. This method detects a rearranged IgH gene from a single B lymphocyte which may or may not represent the neoplastic B-cell population. We therefore performed multiple snPCR (three to five) experiments on the same CSF sample, postulating that the detection of a band of the same size and sequence in different PCR runs was highly indicative of a clonal population. 17 consecutive cases with a differential diagnosis of primary (PCNSL) (n=10) or secondary (SCNSL) (n=7) CNS lymphoma or leukaemia were investigated by the new strategy. The clonal nature of the B-cell population was confirmed in 3/10 of suspected PCNSL, and in six other cases the PCR study was indicative of reactive lymphocytosis. One case revealed a clonal B-cell population in the clinical context of an autoimmune disorder. Evidence of clonal B-cell population was found in 4/7 of suspected SCNSL. In one of these cases the detected band and its sequence proved identical to that of the primary nodal lymphoma. We believe that the evaluation of B-cell clonality in CSF requires multiple snPCR amplification on the same sample to compare the size of the products and, if necessary, the DNA sequences to ascertain the diagnosis of malignancy in equivocal cytologic and clinical findings.
Galoin
S
S
Department of Pathology and CIGH/CNRS, CHU-Purpan, Toulouse, France.
Daste
G
G
Apoil
P A
PA
Chollet
F
F
Roda
D
D
Blancher
A
A
Delsol
G
G
Chittal
S
S
al Saati
T
T
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D005260:Female; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D015448:Leukemia, B-Cell / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D008218:Lymphocytosis / Q000134:cerebrospinal fluid; D016393:Lymphoma, B-Cell / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D008297:Male; D008577:Meningeal Neoplasms / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D007143:Immunoglobulin Heavy Chains