Graduate School of Medicine and Veterinary Medicine, University of Miyazaki, Miyazaki, Japan.
Faculty of Veterinary Medicine, Benha University, Toukh, Egypt.
Transbound Emerg Dis. 2020 Jul;67(4):1671-1676. doi: 10.1111/tbed.13506. Epub 2020 Feb 17.
Enzootic bovine leucosis (EBL) is a neoplastic disease of cattle caused by Bovine leukaemia virus (BLV). EBL causes great economic losses, so a fast and reliable diagnostic method is critical for understanding the status of BLV. This will allow us to control BLV infections efficiently and mitigate economic losses. In this study, we established a direct diagnostic test for BLV using dried blood-spotted filter papers without sample pre-treatment. The study was based on 159 clinical blood specimens collected in EDTA from one farm in Kyushu, Japan. The blood-spotted filter papers were used as the template for direct filter PCR. When an ELISA was used as the diagnostic gold standard, the sensitivity and specificity of the direct filter PCR were 90.1% and 97.5%, respectively. The kappa value for the direct filter PCR and real-time PCR methods was 0.97. The dried blood samples spotted onto filter papers were stable for at least 10 days at room temperature, even when the samples were from cattle with a low BLV proviral load. Direct filter PCR is a rapid, easy, reliable and cost-effective diagnostic test that directly detects the BLV proviral genome in clinical blood specimens without DNA extraction. Moreover, it simplifies the collection, transportation and storage procedures for clinical blood specimens.
牛白血病（EBL）是一种由牛白血病病毒（BLV）引起的牛的肿瘤性疾病。EBL 会造成巨大的经济损失，因此，快速可靠的诊断方法对于了解 BLV 的状况至关重要。这将使我们能够有效地控制 BLV 感染并减轻经济损失。在本研究中，我们建立了一种无需样本预处理即可使用干血斑滤纸直接诊断 BLV 的方法。该研究基于从日本九州的一个农场采集的 159 份 EDTA 临床血液样本。血斑滤纸用作直接滤过 PCR 的模板。当 ELISA 用作诊断金标准时，直接滤过 PCR 的灵敏度和特异性分别为 90.1%和 97.5%。直接滤过 PCR 和实时 PCR 方法的 Kappa 值为 0.97。干血斑滤纸在室温下至少稳定 10 天，即使样本来自 BLV 前病毒载量低的牛。直接滤过 PCR 是一种快速、简便、可靠且具有成本效益的诊断检测方法，无需 DNA 提取即可直接检测临床血液样本中的 BLV 前病毒基因组。此外，它简化了临床血液样本的采集、运输和储存程序。
