Nonradioactive detection of monoclonal immunoglobulin heavy chain gene rearrangement with PCR-SSCP. 1995

U Thunberg, and M Alemi, and C Sundström, and J Sällström
Department of Pathology, University of Uppsala, Sweden.

The rearrangement of the immunoglobulin heavy chain gene can be used as a marker of B-cell lineage and clonality. By using polymerase chain reaction (PCR) with variable-region and joining-region specific primers it is possible to detect the rearrangement of a small amount of clonal B cells, as described by several groups. The specific PCR product can be detected after amplification with gel electrophoresis and ethidium bromide staining. The DNA fragments obtained from different clones are, however, approximately of the same size, making it difficult to distinguish between the clones by simple electrophoresis and ethidium bromide staining, as described in many reports. Single-strand conformational polymorphism (SSCP) was evaluated as a method to detect specific clonal amplicons in a mixture of PCR-amplified products. Unique patterns were obtained for different B-cell clones, detectable in mixtures of 0.25% clonal cells in normal cells. It is concluded that SSCP is a valuable method for the specificity control of PCR in B-lymphocyte clonality analyses. The advantages of the described method over previously published techniques are increased specificity, simplicity without radioactivity, and rapidity.

UI MeSH Term Description Entries
D007143 Immunoglobulin Heavy Chains The largest of polypeptide chains comprising immunoglobulins. They contain 450 to 600 amino acid residues per chain, and have molecular weights of 51-72 kDa. Immunoglobulins, Heavy-Chain,Heavy-Chain Immunoglobulins,Ig Heavy Chains,Immunoglobulin Heavy Chain,Immunoglobulin Heavy Chain Subgroup VH-I,Immunoglobulin Heavy Chain Subgroup VH-III,Heavy Chain Immunoglobulins,Heavy Chain, Immunoglobulin,Heavy Chains, Ig,Heavy Chains, Immunoglobulin,Immunoglobulin Heavy Chain Subgroup VH I,Immunoglobulin Heavy Chain Subgroup VH III,Immunoglobulins, Heavy Chain
D002051 Burkitt Lymphoma A form of undifferentiated malignant LYMPHOMA usually found in central Africa, but also reported in other parts of the world. It is commonly manifested as a large osteolytic lesion in the jaw or as an abdominal mass. B-cell antigens are expressed on the immature cells that make up the tumor in virtually all cases of Burkitt lymphoma. The Epstein-Barr virus (HERPESVIRUS 4, HUMAN) has been isolated from Burkitt lymphoma cases in Africa and it is implicated as the causative agent in these cases; however, most non-African cases are EBV-negative. African Lymphoma,Burkitt Cell Leukemia,Burkitt Tumor,Lymphoma, Burkitt,Burkitt Leukemia,Burkitt's Leukemia,Burkitt's Lymphoma,Burkitt's Tumor,Leukemia, Lymphoblastic, Burkitt-Type,Leukemia, Lymphocytic, L3,Lymphocytic Leukemia, L3,Burkitts Leukemia,Burkitts Lymphoma,Burkitts Tumor,L3 Lymphocytic Leukemia,L3 Lymphocytic Leukemias,Leukemia, Burkitt,Leukemia, Burkitt Cell,Leukemia, Burkitt's,Leukemia, L3 Lymphocytic,Lymphoma, African,Lymphoma, Burkitt's,Tumor, Burkitt,Tumor, Burkitt's
D002648 Child A person 6 to 12 years of age. An individual 2 to 5 years old is CHILD, PRESCHOOL. Children
D002999 Clone Cells A group of genetically identical cells all descended from a single common ancestral cell by mitosis in eukaryotes or by binary fission in prokaryotes. Clone cells also include populations of recombinant DNA molecules all carrying the same inserted sequence. (From King & Stansfield, Dictionary of Genetics, 4th ed) Clones,Cell, Clone,Cells, Clone,Clone,Clone Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000328 Adult A person having attained full growth or maturity. Adults are of 19 through 44 years of age. For a person between 19 and 24 years of age, YOUNG ADULT is available. Adults
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D015322 Gene Rearrangement, B-Lymphocyte Ordered rearrangement of B-lymphocyte variable gene regions coding for the IMMUNOGLOBULIN CHAINS, thereby contributing to antibody diversity. It occurs during the differentiation of the IMMATURE B-LYMPHOCYTES. B-Cell Gene Rearrangement,B-Lymphocyte Gene Rearrangement,Gene Rearrangement, B-Cell,B Cell Gene Rearrangement,B Lymphocyte Gene Rearrangement,B-Cell Gene Rearrangements,B-Lymphocyte Gene Rearrangements,Gene Rearrangement, B Cell,Gene Rearrangement, B Lymphocyte,Gene Rearrangements, B-Cell,Gene Rearrangements, B-Lymphocyte,Rearrangement, B-Cell Gene,Rearrangement, B-Lymphocyte Gene,Rearrangements, B-Cell Gene,Rearrangements, B-Lymphocyte Gene
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain
D016393 Lymphoma, B-Cell A group of heterogeneous lymphoid tumors generally expressing one or more B-cell antigens or representing malignant transformations of B-lymphocytes. B-Cell Lymphoma,B Cell Lymphoma,B-Cell Lymphomas,Lymphoma, B Cell,Lymphomas, B-Cell

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