Several methods have been developed for the detection of minimal residual disease (MRD) in B cell tumors. Chromosomal translocations or the rearrangement of the immunoglobulin heavy chain (IgH) and T cell receptor genes are generally employed. We report a novel PCR method to detect MRD using IgH genes. IgH rearranged variable region (VDJ) were amplified from tumor specimens using consensus primers for variable and joining region genes. Complementarity-determining regions (CDR) were identified and used to generate tumor-specific primers. Two-round amplifications using primers derived from CDRs and joining or constant regions were performed for MRD detection. IgH nested-PCR approach was tested on a panel of 75 B cell tumors including acute lymphoblastic and chronic lymphocytic leukemias, non-Hodgkin's lymphomas and multiple myelomas. A VDJ sequence was obtained in 62 out of 75 cases (83%). Sensitivity using DNA or cDNA templates was 10(-5) and (-6), respectively. This method is specific and sensitive and provides a simple, non-radioactive approach for the evaluation of MRD in B cell tumors.

A novel nested-PCR strategy for the detection of rearranged immunoglobulin heavy-chain genes in B cell tumors / C. Voena, M. Ladetto, M. Astolfi, D. Provan, J. G. Gribben, M. Boccadoro, A. Pileri, P. Corradini. - In: LEUKEMIA. - ISSN 0887-6924. - 11:10(1997 Oct), pp. 1793-1798.

A novel nested-PCR strategy for the detection of rearranged immunoglobulin heavy-chain genes in B cell tumors

P. Corradini
Ultimo
1997

Abstract

Several methods have been developed for the detection of minimal residual disease (MRD) in B cell tumors. Chromosomal translocations or the rearrangement of the immunoglobulin heavy chain (IgH) and T cell receptor genes are generally employed. We report a novel PCR method to detect MRD using IgH genes. IgH rearranged variable region (VDJ) were amplified from tumor specimens using consensus primers for variable and joining region genes. Complementarity-determining regions (CDR) were identified and used to generate tumor-specific primers. Two-round amplifications using primers derived from CDRs and joining or constant regions were performed for MRD detection. IgH nested-PCR approach was tested on a panel of 75 B cell tumors including acute lymphoblastic and chronic lymphocytic leukemias, non-Hodgkin's lymphomas and multiple myelomas. A VDJ sequence was obtained in 62 out of 75 cases (83%). Sensitivity using DNA or cDNA templates was 10(-5) and (-6), respectively. This method is specific and sensitive and provides a simple, non-radioactive approach for the evaluation of MRD in B cell tumors.
Neoplasm, Residual; Sensitivity and Specificity; Humans; Gene Rearrangement, B-Lymphocyte, Heavy Chain; Genes, Immunoglobulin; DNA, Neoplasm; Gene Amplification; Multiple Myeloma; Lymphoma, B-Cell; Polymerase Chain Reaction; Leukemia, Lymphocytic, Chronic, B-Cell; Burkitt Lymphoma; DNA Primers
Settore MED/15 - Malattie del Sangue
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/181127
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