Direct polymerase chain reaction from blood and tissue samples for rapid diagnosis of bovine leukemia virus infection

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Author(s)

    • NISHIMORI Asami NISHIMORI Asami
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan
    • KONNAI Satoru KONNAI Satoru
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan
    • IKEBUCHI Ryoyo IKEBUCHI Ryoyo
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan
    • OKAGAWA Tomohiro OKAGAWA Tomohiro
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan
    • NAKAHARA Ayako NAKAHARA Ayako
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan
    • MURATA Shiro MURATA Shiro
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan
    • OHASHI Kazuhiko OHASHI Kazuhiko
    • Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita-ku Kita 18- jo Nishi 9-chome, Sapporo, Hokkaido 060–0818, Japan

Abstract

Bovine leukemia virus (BLV) infection induces bovine leukemia in cattle and causes significant financial harm to farmers and farm management. There is no effective therapy or vaccine; thus, the diagnosis and elimination of BLV-infected cattle are the most effective method to eradicate the infection. Clinical veterinarians need a simpler and more rapid method of diagnosing infection, because both nested polymerase chain reaction (PCR) and real-time PCR are labor intensive, time-consuming, and require specialized molecular biology techniques and expensive equipment. In this study, we describe a novel PCR method for amplifying the BLV provirus from whole blood, thus eliminating the need for DNA extraction. Although the sensitivity of PCR directly from whole blood (PCR-DB) samples as measured in bovine blood containing BLV-infected cell lines was lower than that of nested PCR, the PCR-DB technique showed high specificity and reproducibility. Among 225 clinical samples, 49 samples were positive by nested PCR, and 37 samples were positive by PCR-DB. There were no false positive samples; thus, PCR-DB sensitivity and specificity were 75.51% and 100%, respectively. However, the provirus loads of the samples detected by nested PCR and not PCR-DB were quite low. Moreover, PCR-DB also stably amplified the BLV provirus from tumor tissue samples. PCR-DB method exhibited good reproducibility and excellent specificity and is suitable for screening of thousands of cattle, thus serving as a viable alternative to nested PCR and real-time PCR.

Journal

  • Journal of Veterinary Medical Science

    Journal of Veterinary Medical Science 78(5), 791-796, 2016

    JAPANESE SOCIETY OF VETERINARY SCIENCE

Codes

  • NII Article ID (NAID)
    130005154280
  • NII NACSIS-CAT ID (NCID)
    AA10796138
  • Text Lang
    ENG
  • ISSN
    0916-7250
  • NDL Article ID
    027449715
  • NDL Call No.
    Z18-350
  • Data Source
    NDL  J-STAGE 
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