Fluorescence in situ hybridization analysis of immunoglobulin heavy chain translocations in plasma cell myeloma using intact paraffin sections and simultaneous CD138 immunofluorescence.
Fluorescence in situ hybridization (FISH) studies are much more sensitive than classical cytogenetics for identification of karyotypic abnormalities in plasma cell myeloma. However, FISH analysis of bone marrow samples is often challenging because of a large number of admixed non-neoplastic hematopoietic elements. In this report, we describe a novel method using FISH analysis of intact paraffin sections of formalin-fixed, bone marrow clot preparations with simultaneous CD138 tyramine signal amplification (TSA)-mediated immunofluorescence. We studied 22 cases of plasma cell myeloma for translocations involving the immunoglobulin heavy chain locus that are of known diagnostic and/or prognostic significance. All cases were analyzed using dual color, break-apart immunoglobulin heavy chain probe and dual color, dual fusion probes for t(11;14)(q13;q32) and t(4;14)(p16;q32). TSA-mediated fluorochrome deposition in CD138+ cells was unaltered by protease pretreatment. Translocations were identified in 10 cases, including five with t(11;14)(q13;q32) and three with t(4;14)(p16.3;q32). When present, abnormalities were identified in a large percentage of CD138+ cells (47 to 93%, median 84%). This technique allows for efficient molecular cytogenetic analysis of plasma cell myeloma using routinely archived paraffin-embedded material.
Cook
James R
JR
Department of Clinical Pathology, L11, 9500 Euclid Ave., Cleveland, OH 44195, USA. cookj2@ccf.org
Hartke
Marybeth
M
Pettay
James
J
Tubbs
Raymond R
RR
D002880:Chromosomes, Human, Pair 11; D002883:Chromosomes, Human, Pair 14; D020732:Cytogenetic Analysis; D005455:Fluorescent Antibody Technique; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008562:Membrane Glycoproteins / Q000032:analysis*; D009101:Multiple Myeloma / Q000235:genetics*; D016612:Paraffin Embedding; D011509:Proteoglycans / Q000032:analysis*; D053668:Syndecan-1; D053667:Syndecans; D014178:Translocation, Genetic
D007143:Immunoglobulin Heavy Chains; D008562:Membrane Glycoproteins; D011509:Proteoglycans; C508241:SDC1 protein, human; D053668:Syndecan-1; D053667:Syndecans