Okagawa Tomohiro, Nojiri Naomi, Yoshida-Furihata Hazuka, Nao Naganori, Tominaga Misono, Kohara Junko, Gondaira Satoshi, Higuchi Hidetoshi, Takeda Yohei, Ogawa Haruko, Yamada Shinji, Murakami Kenji, Suzuki Yasunori, Takai Shinji, Maezawa Masaki, Inokuma Hisashi, Shimizu Kaori, Inoshima Yasuo, Usui Tatsufumi, Tagawa Michihito, Yamamoto Mari, Mekata Hirohisa, Esaki Mana, Ozawa Makoto, Matsudaira Takahiro, Maekawa Naoya, Murata Shiro, Ohashi Kazuhiko, Saito Masumichi, Konnai Satoru
Department of Advanced Pharmaceutics, Faculty of Veterinary Medicine, Hokkaido University, Hokkaido, Japan.
Center for Emergency Preparedness and Response, National Institute of Infectious Diseases, Tokyo, Japan.
J Vet Med Sci. 2025 May 15;87(5):551-558. doi: 10.1292/jvms.25-0031. Epub 2025 Mar 28.
Bovine leukemia virus (BLV), a retrovirus that is widespread worldwide, causes enzootic bovine leukosis (EBL), a B-cell leukemia/lymphoma with a poor prognosis that ultimately results in death. In Japan, the number of cattle infected with this virus is increasing, and it is estimated more than 35% of cattle are currently infected. Since no vaccines or treatments against BLV infection are currently available, it is important to establish a method of early diagnosis for EBL to reduce economic losses caused by the disposal of EBL cattle in Japan, where a large number of expensive beef cattle are raised. We previously developed Rapid Amplification of the Integration Site without Interference by Genomic DNA Contamination (RAISING), a cost-effective, rapid, and sensitive method for the clonality analysis of BLV-infected cells. Despite its usefulness for the early diagnosis of EBL, RAISING had drawbacks preventing its practical application. Here, we report the development of an improved method, RAISING ver.2, and its performance. Compared to BLV clonality analysis using the previous method, RAISING ver.2 was found to maintain high accuracy and reproducibility despite its simplification. Moreover, its performance was also validated in a multicenter validation study. Taken together, our results strongly suggest that RAISING ver.2 can be fully utilized in clinical practice. Successful commercialization of a RAISING test kit could overcome the concerns of livestock farmers suffering from EBL, thereby promoting a stable supply of Japanese beef, both domestically and internationally.
牛白血病病毒（BLV）是一种在全球广泛传播的逆转录病毒，可引发地方流行性牛白血病（EBL），这是一种预后不良的B细胞白血病/淋巴瘤，最终会导致死亡。在日本，感染这种病毒的牛的数量正在增加，据估计目前超过35%的牛受到感染。由于目前尚无针对BLV感染的疫苗或治疗方法，因此建立一种EBL早期诊断方法对于减少日本因处理EBL病牛而造成的经济损失非常重要，因为日本饲养了大量昂贵的肉牛。我们之前开发了一种不受基因组DNA污染干扰的整合位点快速扩增方法（RAISING），这是一种用于BLV感染细胞克隆性分析的经济高效、快速且灵敏的方法。尽管RAISING对EBL的早期诊断很有用，但它存在一些缺点，阻碍了其实际应用。在此，我们报告一种改进方法RAISING ver.2的开发及其性能。与使用先前方法进行的BLV克隆性分析相比，尽管RAISING ver.2进行了简化，但仍保持了高准确性和可重复性。此外，其性能也在一项多中心验证研究中得到了验证。综上所述，我们的结果强烈表明RAISING ver.2可在临床实践中充分利用。成功将RAISING检测试剂盒商业化可以消除患有EBL的养殖户的担忧，从而促进日本牛肉在国内和国际上的稳定供应。
