title: Construction of a multicolor GeneScan analytical system to detect clonal rearrangements of immunoglobulin and T cell receptor genes in canine lymphoid tumors.
abstract: Polymerase chain reaction (PCR) amplification to detect immunoglobulin heavy chain (IgH) and T cell receptor γ-chain (TCRγ) gene rearrangements has recently become widely used as part of the diagnostic strategy for lymphoid tumors in dogs. In this study, we constructed a multicolor GeneScan analytical system to improve the sensitivity and resolution of the clonality analysis of antigen receptor gene rearrangements in dogs. We used 7 reactions per sample, with 2 PCR conditions, to amplify IgH/TCRγ and control genes. By using multicolor-labeled primers, these 7 PCR products could be combined into 3 tubes before capillary electrophoresis. Clonal rearrangement of the IgH/TCRγ genes was detected in 93.3% of dogs with multicentric lymphoma and 84.6% of dogs with gastrointestinal lymphoma. Detection sensitivity of the clonally expanded cells in the background of normal peripheral blood mononuclear cells was 1-10%. The multicolor GeneScan analytical system developed here may prove to be helpful for the diagnosis of lymphoid tumors in dogs.
authors.affiliation: Molecular Diagnostic Laboratory, Veterinary Medical Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Electronic address: ayukogk@mail.ecc.u-tokyo.ac.jp.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Goto-Koshino
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mochizuki
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Sato
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Ko
authors.initials: K
authors.lastname: Nakashima
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Saaya
authors.initials: S
authors.lastname: Hiyoshi
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Aki
authors.initials: A
authors.lastname: Fujiwara-Igarashi
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Maeda
authors.affiliation: Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Nakamura
authors.affiliation: Department of Veterinary Pathology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Fujino
authors.affiliation: Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Ohno
authors.affiliation: Molecular Diagnostic Laboratory, Veterinary Medical Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Department of Veterinary Internal Medicine Graduate, School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Tsujimoto
mesh_terms: D000818:Animals; D004283:Dog Diseases / Q000235:genetics* / Q000276:immunology; D004285:Dogs / Q000235:genetics; D015321:Gene Rearrangement / Q000235:genetics*; D019675:Genes, T-Cell Receptor gamma / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007136:Immunoglobulins / Q000235:genetics*; D008223:Lymphoma / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*
chemical_list: D007143:Immunoglobulin Heavy Chains; D007136:Immunoglobulins; D011948:Receptors, Antigen, T-Cell
keywords: Dog; GeneScan; Multicolor; PCR; lympHoma