A NOVEL NESTED-PCR STRATEGY FOR THE DETECTION OF REARRANGED IMMUNOGLOBULIN HEAVY-CHAIN GENES IN B-CELL TUMORS

Several methods have been developed for the detection of minimal resid ual disease (MRD) in B cell tumors. Chromosomal translocations or the rearrangement of the immunoglobulin heavy chain (IgH) and T cell recep tor genes are generally employed. We report a novel PCR method to dete ct MRD using IgH genes. IgH rearranged variable region (VDJ) were ampl ified from tumor specimens using consensus primers for variable and jo ining region genes. Complementarity-determining regions (CDR) were ide ntified and used to generate tumor-specific primers. Two-round amplifi cations using primers derived from CDRs and joining or constant region s were performed for MRD detection. IgH nested-PCR approach was tested on a panel of 75 B cell tumors including acute lymphoblastic and chro nic lymphocytic leukemias, non-Hodgkin's lymphomas and multiple myelom as. A VDJ sequence was obtained in 62 out of 75 cases (83%). Sensitivi ty using DNA or cDNA templates was 10(-5) and (-6), respectively. This method is specific and sensitive and provides a simple, non-radioacti ve approach for the evaluation of MRD in B cell tumors.