Development of a microchip electrophoresis-based, high-throughput PCR-RFLP method to type Tax 233 variants of bovine leu
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BRIEF REPORT
Development of a microchip electrophoresis‑based, high‑throughput PCR‑RFLP method to type Tax 233 variants of bovine leukemia virus in Japan Chihiro Ochiai1 · Yuzuru Katagiri1 · Sota Kobayashi2 · Ikunori Naitoh3 · Syuji Yoneyama4 · Keisuke Tomita4 · Leng Dongze4 · Hirokazu Hikono5 · Kenji Murakami1,3,4 Received: 2 July 2020 / Accepted: 4 September 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). We used microchip electrophoresis in combination with automatic image analysis to develop a novel high-throughput PCR–RFLP to type the gene sequences that encode BLV Tax 233. This method revealed that 233L-Tax is more prevalent than 233P-Tax in cattle in Japan. The proportion infected with BLV carrying the gene encoding 233L-Tax was significantly higher in Holstein cattle than in Japanese Black cattle. Holsteins infected with BLV encoding 233L-Tax had higher proviral loads than did Holsteins infected with BLV encoding 233P-Tax and Japanese Blacks infected with BLV encoding 233L-Tax or 233P-Tax. The novel method developed in this study will be a useful tool for identifying cattle harboring BLV with a higher risk of EBL and viral transmission. Bovine leukemia virus (BLV) is an RNA virus belonging to the genus Deltaretrovirus, family Retroviridae; it is a close relative of human T cell leukemia virus types 1 and 2 (HTLV-1 and 2) [1]. BLV is the causative agent of enzootic bovine leukosis (EBL), a malignant B cell lymphoma. The retroviral genome uses its own integrase to integrate itself into the host genome, where it becomes a provirus and persists throughout life. Many cattle are asymptomatic carriers. Handling Editor: Zhongjie Shi. * Kenji Murakami muraken@iwate‑u.ac.jp 1
Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, 3‑18‑8 Ueda, Morioka, Iwate 020‑8550, Japan
2
Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research Organization, 3‑1‑5 Kannondai, Tsukuba, Ibaraki 305‑0856, Japan
3
The United Graduate School of Veterinary Sciences, Gifu University, 1‑1 Yanagido, Gifu City, Gifu 501‑1193, Japan
4
Graduate School of Veterinary Sciences, Iwate University, 3‑18‑8 Ueda, Morioka, Iwate 020‑8550, Japan
5
Department of Animal Sciences, Teikyo University of Science, 2‑2‑1 Senju‑sakuragi, Adachi‑ku, Tokyo 120‑0045, Japan
Persistent lymphocytosis (PL) occurs in approximately 30% of BLV-infected cattle and 0.1% to 5% of those that develop EBL [2, 3]. In Japan, a nationwide survey of BLV revealed that the prevalence was 28.7% in beef breeding cattle and 40.7% in dairy cattle [4]. The annual number of EBL outbreaks in Japan was reported to be only 99 in 1998; however, this has rapidly increased, with 4113 outbreaks in 2019 [5]. EBL is a notifiable disease by the Act on Domestic Animal Infectious Diseases Control, and if EBL is found upon meat inspection, the entire carcass must be discard
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