title: Standardization of Quantitative PCR for Human T-Cell Leukemia Virus Type 1 in Japan: a Collaborative Study.
abstract: Quantitative PCR (qPCR) analysis of human T-cell leukemia virus type 1 (HTLV-1) was used to assess the amount of HTLV-1 provirus DNA integrated into the genomic DNA of host blood cells. Accumulating evidence indicates that a high proviral load is one of the risk factors for the development of adult T-cell leukemia/lymphoma and HTLV-1-associated myelopathy/tropical spastic paraparesis. However, interlaboratory variability in qPCR results makes it difficult to assess the differences in reported proviral loads between laboratories. To remedy this situation, we attempted to minimize discrepancies between laboratories through standardization of HTLV-1 qPCR in a collaborative study. TL-Om1 cells that harbor the HTLV-1 provirus were serially diluted with peripheral blood mononuclear cells to prepare a candidate standard. By statistically evaluating the proviral loads of the standard and those determined using in-house qPCR methods at each laboratory, we determined the relative ratios of the measured values in the laboratories to the theoretical values of the TL-Om1 standard. The relative ratios of the laboratories ranged from 0.84 to 4.45. Next, we corrected the proviral loads of the clinical samples from HTLV-1 carriers using the relative ratio. As expected, the overall differences between the laboratories were reduced by half, from 7.4-fold to 3.8-fold on average, after applying the correction. HTLV-1 qPCR can be standardized using TL-Om1 cells as a standard and by determining the relative ratio of the measured to the theoretical standard values in each laboratory.
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Madoka
authors: http://orcid.org/0000-0002-3064-5219
authors: M
authors: Kuramitsu
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kazu
authors: K
authors: Okuma
authors: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors: Tadanori
authors: T
authors: Yamochi
authors: Department of Rare Diseases Research, Institute of Medical Science, St. Marianna University School of Medicine, Kawasaki, Japan.
authors: Tomoo
authors: T
authors: Sato
authors: Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors: Daisuke
authors: D
authors: Sasaki
authors: Department of Laboratory Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors: Hiroo
authors: H
authors: Hasegawa
authors: Department of Rheumatology, Infectious Diseases and Laboratory Medicine, University of Miyazaki, Miyazaki, Japan.
authors: Kazumi
authors: K
authors: Umeki
authors: Division of Molecular Pathology, Center for Chronic Viral Diseases, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.
authors: Ryuji
authors: R
authors: Kubota
authors: Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
authors: Rieko
authors: R
authors: Sobata
authors: Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
authors: Chieko
authors: C
authors: Matsumoto
authors: Department of Special Testing, SRL Inc., Tokyo, Japan.
authors: Noriaki
authors: N
authors: Kaneko
authors: Department of Special Testing, SRL Inc., Tokyo, Japan.
authors: Isao
authors: I
authors: Naruse
authors: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors: Makoto
authors: M
authors: Yamagishi
authors: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors: Makoto
authors: M
authors: Nakashima
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Haruka
authors: H
authors: Momose
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kumiko
authors: K
authors: Araki
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Takuo
authors: T
authors: Mizukami
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Saeko
authors: S
authors: Mizusawa
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Yoshiaki
authors: Y
authors: Okada
authors: Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Masaki
authors: M
authors: Ochiai
authors: Department of Hematology, Imamura Bun-in Hospital, Kagoshima, Japan.
authors: Atae
authors: A
authors: Utsunomiya
authors: Department of Hematology, Osaka General Hospital of West Japan Railway Company, Osaka, Japan.
authors: Ki-Ryang
authors: KR
authors: Koh
authors: Blood Transfusion Center, Oita University Faculty of Medicine, Oita, Japan.
authors: Masao
authors: M
authors: Ogata
authors: Department of Hematology, Kumamoto University of Medicine, Kumamoto, Japan.
authors: Kisato
authors: K
authors: Nosaka
authors: Department of Hematology/Oncology, Research Hospital, Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors: Kaoru
authors: K
authors: Uchimaru
authors: Department of Frontier Life Science, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors: Masako
authors: M
authors: Iwanaga
authors: Japanese Red Cross Kyushu Block Blood Center, Fukuoka, Japan.
authors: Yasuko
authors: Y
authors: Sagara
authors: Department of Rare Diseases Research, Institute of Medical Science, St. Marianna University School of Medicine, Kawasaki, Japan.
authors: Yoshihisa
authors: Y
authors: Yamano
authors: Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
authors: Masahiro
authors: M
authors: Satake
authors: Department of Rheumatology, Infectious Diseases and Laboratory Medicine, University of Miyazaki, Miyazaki, Japan.
authors: Akihiko
authors: A
authors: Okayama
authors: Department of Ophthalmology and Visual Science, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors: Manabu
authors: M
authors: Mochizuki
authors: Division of Molecular Pathology, Center for Chronic Viral Diseases, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.
authors: Shuji
authors: S
authors: Izumo
authors: Department of Obstetrics and Gynecology, University of Toyama, Toyama, Japan.
authors: Shigeru
authors: S
authors: Saito
authors: Department of Pediatrics, Showa University School of Medicine, Tokyo, Japan.
authors: Kazuo
authors: K
authors: Itabashi
authors: Nagasaki Harbor Medical Center, ITREC, Nagasaki, Japan.
authors: Shimeru
authors: S
authors: Kamihira
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kazunari
authors: K
authors: Yamaguchi
authors: Department of Medical Genome Sciences, Laboratory of Tumor Cell Biology, Graduate School of Frontier Sciences, The University of Tokyo, Tokyo, Japan.
authors: Toshiki
authors: T
authors: Watanabe
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan 130hama@niid.go.jp.
authors: Isao
authors: I
authors: Hamaguchi
mesh_terms: D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D015490:HTLV-I Infections / Q000235:genetics / Q000821:virology; D015368:Human T-lymphotropic virus 1 / Q000235:genetics*; D006801:Humans; D007564:Japan; D019169:Jurkat Cells; D015458:Leukemia, T-Cell / Q000235:genetics / Q000821:virology; D007963:Leukocytes, Mononuclear / Q000821:virology; D011533:Proviruses / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D019562:Viral Load / Q000235:genetics*; D016662:Virus Integration / Q000235:genetics
chemical_list: D004279:DNA, Viral