Ullah Shakir, Notsu Kosuke, Saito Akatsuki, Okabayashi Tamaki, Mekata Hirohisa, Isoda Norikazu, Sekiguchi Satoshi
Graduate School of Medicine and Veterinary Medicine, University of Miyazaki, Miyazaki, 889-1692, Japan.
Livestock and Dairy Development Department Balochistan, Quetta, 87300, Pakistan.
Arch Virol. 2024 Dec 12;170(1):15. doi: 10.1007/s00705-024-06207-z.
Bovine viral diarrhea (BVD), caused by bovine viral diarrhea virus (BVDV), has a significant economic impact on affected farms worldwide. For effective disease control, it is crucial to select an appropriate vaccine based on the specific genotype of BVDV. Therefore, developing a rapid and reliable assay to detect and genotype BVDV is imperative for controlling the spread of disease. In this study, we developed a TaqMan assay to detect and genotype BVDV types 1 and 2 directly in bovine serum without extraction of RNA. The direct BVDV TaqMan assay effectively detected both BVDV1 and BVDV2 with confirmed specificity and showed no cross-reactivity with any of the other viruses tested, including bovine respiratory syncytial virus, bovine coronavirus, Akabane virus, bovine herpesvirus 1, bovine parainfluenza virus 3, bovine immunodeficiency virus, and bovine leukemia virus. The assay could detect the virus in serum samples with a titer as low as 10 TCID/mL in two out of three trials for BVDV1 and all three trials for BVDV2, indicating that its sensitivity is equivalent to that of virus isolation. Our findings represent a significant advancement in BVDV detection and typing directly from bovine serum.
牛病毒性腹泻（BVD）由牛病毒性腹泻病毒（BVDV）引起，对全球受影响的养殖场具有重大经济影响。为了有效控制疾病，根据BVDV的特定基因型选择合适的疫苗至关重要。因此，开发一种快速可靠的检测方法来检测BVDV并进行基因分型对于控制疾病传播势在必行。在本研究中，我们开发了一种TaqMan检测方法，可直接在牛血清中检测BVDV 1型和2型并进行基因分型，无需提取RNA。直接BVDV TaqMan检测方法有效地检测了BVDV1和BVDV2，特异性得到确认，并且与所测试的任何其他病毒均无交叉反应，包括牛呼吸道合胞病毒、牛冠状病毒、赤羽病病毒、牛疱疹病毒1型、牛副流感病毒3型、牛免疫缺陷病毒和牛白血病病毒。该检测方法在三分之二的BVDV1试验和所有BVDV2试验中能够检测到滴度低至10 TCID/mL的血清样本中的病毒，表明其灵敏度与病毒分离相当。我们的研究结果代表了直接从牛血清中检测和分型BVDV的重大进展。
