Validation of sixteen leukemia and lymphoma cell lines as controls for molecular gene rearrangement assays.
BACKGROUND
Assays for rearrangement of the immunoglobulin, T-cell receptor, bcr/abl, and bcl-2 genes are valuable tools to aid in the diagnosis of leukemias and lymphomas and are now offered by many pathology laboratories. However, there is a lack of well-characterized and validated calibrators and positive controls for these assays. We therefore evaluated 16 readily available leukemia and lymphoma cell lines for their potential use as controls.


METHODS
DNA and RNA were isolated from each cell line and analyzed by Southern blot and PCR or reverse transcription-PCR (RT-PCR). Rearrangements in the IgJ(H), IgJ(kappa), TcR-beta or TcR-gamma, bcr/abl, and bcl-2 genes were detected by commercially available probes and primers. Cell lineages were confirmed by immunophenotyping.


RESULTS
Immunoglobulin and T-cell receptor gene rearrangements were identified in five B- and three T-cell lines, respectively. Two cell lines tested positive for the bcr/abl gene, and one was positive for the bcl-2 gene rearrangement by Southern blot.


CONCLUSIONS
The 16 cell lines studied can be used as positive controls in molecular detection assays for gene rearrangements. The parallel processing of these cell lines with clinical samples can serve to quality control the experimental procedures from the first step of DNA or RNA extraction to the final step of result analysis.
Yao
Rong
R
Wadsworth Center, New York State Department of Health, Empire State Plaza, Albany 12201, USA.
Rich
Steven A
SA
Schneider
Erasmus
E
D001402:B-Lymphocytes / Q000378:metabolism; D015139:Blotting, Southern; D005434:Flow Cytometry; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D015322:Gene Rearrangement, B-Lymphocyte; D015329:Gene Rearrangement, T-Lymphocyte; D019673:Genes, T-Cell Receptor alpha; D019675:Genes, T-Cell Receptor gamma; D019254:Genes, bcl-2; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D016130:Immunophenotyping; D007938:Leukemia / Q000473:pathology*; D008223:Lymphoma / Q000473:pathology*; D016133:Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000378:metabolism; D014407:Tumor Cells, Cultured
D007136:Immunoglobulins; D016044:Fusion Proteins, bcr-abl