title: Identification of TL-Om1, an adult T-cell leukemia (ATL) cell line, as reference material for quantitative PCR for human T-lymphotropic virus 1.
abstract: Quantitative PCR (qPCR) for human T-lymphotropic virus 1 (HTLV-1) is useful for measuring the amount of integrated HTLV-1 proviral DNA in peripheral blood mononuclear cells. Many laboratories in Japan have developed different HTLV-1 qPCR methods. However, when six independent laboratories analyzed the proviral load of the same samples, there was a 5-fold difference in their results. To standardize HTLV-1 qPCR, preparation of a well-defined reference material is needed. We analyzed the integrated HTLV-1 genome and the internal control (IC) genes of TL-Om1, a cell line derived from adult T-cell leukemia, to confirm its suitability as a reference material for HTLV-1 qPCR. Fluorescent in situ hybridization (FISH) showed that HTLV-1 provirus was monoclonally integrated in chromosome 1 at the site of 1p13 in the TL-Om1 genome. HTLV-1 proviral genome was not transferred from TL-Om1 to an uninfected T-cell line, suggesting that the HTLV-1 proviral copy number in TL-Om1 cells is stable. To determine the copy number of HTLV-1 provirus and IC genes in TL-Om1 cells, we used FISH, digital PCR, and qPCR. HTLV-1 copy numbers obtained by these three methods were similar, suggesting that their results were accurate. Also, the ratio of the copy number of HTLV-1 provirus to one of the IC genes, RNase P, was consistent for all three methods. These findings indicate that TL-Om1 cells are an appropriate reference material for HTLV-1 qPCR.
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Madoka
authors.identifier: http://orcid.org/0000-0002-3064-5219
authors.initials: M
authors.lastname: Kuramitsu
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Kazu
authors.initials: K
authors.lastname: Okuma
authors.affiliation: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Yamagishi
authors.affiliation: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Tadanori
authors.initials: T
authors.lastname: Yamochi
authors.affiliation: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Sanaz
authors.initials: S
authors.lastname: Firouzi
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Haruka
authors.initials: H
authors.lastname: Momose
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Takuo
authors.initials: T
authors.lastname: Mizukami
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Takizawa
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Araki
authors.affiliation: Division of Molecular and Cellular Oncology, Miyagi Cancer Center Research Institute, Natori, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Sugamura
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Kazunari
authors.initials: K
authors.lastname: Yamaguchi
authors.affiliation: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Watanabe
authors.affiliation: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan 130hama@niid.go.jp.
authors.forename: Isao
authors.initials: I
authors.lastname: Hamaguchi
mesh_terms: D045744:Cell Line, Tumor; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007564:Japan; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D019562:Viral Load / Q000379:methods*