Erschienen in:
29.07.2020 | Rapid Communication
A high-throughput detection method for the clonality of Human T-cell leukemia virus type-1-infected cells in vivo
verfasst von:
Masumichi Saito, Hiroo Hasegawa, Shunsuke Yamauchi, So Nakagawa, Daisuke Sasaki, Naganori Nao, Michikazu Tanio, Yusaku Wada, Takahiro Matsudaira, Haruka Momose, Madoka Kuramitsu, Makoto Yamagishi, Makoto Nakashima, Shingo Nakahata, Hidekatsu Iha, Masao Ogata, Yoshitaka Imaizumi, Kaoru Uchimaru, Kazuhiro Morishita, Toshiki Watanabe, Yasushi Miyazaki, Katsunori Yanagihara
Erschienen in:
International Journal of Hematology
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Ausgabe 3/2020
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Abstract
Approximately 10–20 million of Human T-cell leukemia virus type-1 (HTLV-1)-infected carriers have been previously reported, and approximately 5% of these carriers develop adult T-cell leukemia/lymphoma (ATL) with a characteristic poor prognosis. In Japan, Southern blotting has long been routinely performed for detection of clonally expanded ATL cells in vivo, and as a confirmatory diagnostic test for ATL. However, alternative methods to Southern blotting, such as sensitive, quantitative, and rapid analytical methods, are currently required in clinical practice. In this study, we developed a high-throughput method called rapid amplification of integration site (RAIS) that could amplify HTLV-1-integrated fragments within 4 h and detect the integration sites in > 0.16% of infected cells. Furthermore, we established a novel quantification method for HTLV-1 clonality using Sanger sequencing with RAIS products, and the validity of the quantification method was confirmed by comparing it with next-generation sequencing in terms of the clonality. Thus, we believe that RAIS has a high potential for use as an alternative routine molecular confirmatory test for the clonality analysis of HTLV-1-infected cells.