Suhaib Ahmed.
Immunoglobulin Gene rearrangement in Lymphoid malignancies: a simple method for routine use.
Pak J Pathol Jan ;17(3):109-10.

Objective: The study was conducted to optimise and evaluate a PCR based method for detection of clonal Immunoglobulin gene rearrangements in lymphoid malignancies. Design: Prospective analysis of genomic DNA extracted from archival bone marrow smears. Place & Duration: The study was conducted at the Department of Haematology, Armed Forces Institute of Pathology, Rawalpindi between Jun 2000 and Dec 2001. Subjects & Methods: The subjects of the study were 23 cases of Chronic Lymphocytic Leukaemia (CLL) and 40 cases of Acute Lymphoblastic Leukaemia (ALL). All of the cases were diagnosed by the standard morphology and cytochemical methods. Genomic DNA was extracted from the archival unstained bone marrow smears. Polymerase Chain Reaction (PCR) amplification of the IgH gene was done by a VH primer homologous with a highly conserved sequence near the 3` end of FR3 region and a consensus sequence JH primer. The amplified DNA was seen by electrophoresis on 6% polyacrylamide gels. Results: A sharp clonal band ranging from 90-160 bp indicated a positive reaction. Clonal IgH gene rearrangements were seen in 21/23 (91.3%) of ALL and 26/40 (65%) of CLL. Serial dilutions of a DNA sample having IgH gene rearrangement were tested to find out the sensitivity of the assay. The target DNA could be amplified at 1/64 dilution roughly corresponding to <1% malignant cell population.

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