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.
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Acknowledgements
We are grateful to Dr. Masao Matsuoka for providing the ED cell line and a collaborative project of the Joint Study on Prognostic Factors of ATL Development (JSPFAD) for providing DNA obtained from the peripheral blood lymphocytes from HTLV-1 carriers and ATL patients. We also would like to thank Editage (www.editage.com) for English language editing. This work was supported by grants from JSPS KAKENHI (JP17H03594: M.S., JP15K14388: M.S. and M.H., JP15K08647: H.H).
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MS, HH, SY, DS, YW, TM, and MK performed research. SN, NN, MT, and HH performed data analysis. MY, MN, SN, HI, MO, YI, KU, KM, TW, YM, and KY provided patient samples. MS wrote the manuscript. MS and HH supervised the study.
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Saito, M., Hasegawa, H., Yamauchi, S. et al. A high-throughput detection method for the clonality of Human T-cell leukemia virus type-1-infected cells in vivo. Int J Hematol 112, 300–306 (2020). https://doi.org/10.1007/s12185-020-02935-5
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DOI: https://doi.org/10.1007/s12185-020-02935-5