College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China; Key Laboratory of Animal Resistant Biology of Shandong, Ruminant Disease Research Center, College of Life Science, Shandong Normal University, Shandong Province, China.
Key Laboratory of Animal Resistant Biology of Shandong, Ruminant Disease Research Center, College of Life Science, Shandong Normal University, Shandong Province, China.
Mol Cell Probes. 2018 Oct;41:22-26. doi: 10.1016/j.mcp.2018.08.004. Epub 2018 Aug 20.
Bovine respirovirus 3 also known as Bovine parainfluenza virus type 3 (BPIV3) is one of the most important viral respiratory agents of both young and adult cattle. Rapid diagnosis could contribute greatly in containing epidemics and thus avoid economic losses. However, the lack of robust isothermal visual method poses difficulty. In this study, a novel isothermal assay for detecting BPIV3 was established. The method includes a lateral flow dipstick (LFD) assay combined with reverse transcription recombinase polymerase amplification (RT-RPA). First, the analytical sensitivity and specificity of BPIV3 LFD RT-RPA were tested. The LFD RT-RPA assay has a detection limit of up to 100 copies per reaction in 30 min at 38 °C. Then the performance of LFD RT-RPA was evaluated using 95 clinical samples. Compared to qPCR, the LFD RT-RPA assay showed a clinical sensitivity of 94.74%, a clinical specificity of 96.05% and 0.8734 kappa coefficient. These results have demonstrated the efficiency and effectiveness of the method to be developed into a point of care protocol for the diagnosis of BPIV3.
牛呼吸肠道病毒 3 也被称为牛副流感病毒 3 型（BPIV3），是幼牛和成年牛最重要的病毒性呼吸道病原体之一。快速诊断可以极大地有助于控制疫情，从而避免经济损失。然而，缺乏稳健的等温可视化方法是一个难题。在本研究中，建立了一种用于检测 BPIV3 的新型等温测定法。该方法包括侧向流动纸条（LFD）测定法与逆转录重组酶聚合酶扩增（RT-RPA）相结合。首先，测试了 BPIV3 LFD RT-RPA 的分析灵敏度和特异性。LFD RT-RPA 检测法在 38°C 下 30 分钟内的检测限高达 100 个拷贝/反应。然后使用 95 个临床样本评估 LFD RT-RPA 的性能。与 qPCR 相比，LFD RT-RPA 检测法的临床灵敏度为 94.74%，临床特异性为 96.05%，kappa 系数为 0.8734。这些结果表明该方法具有效率和有效性，可开发为用于诊断 BPIV3 的即时护理协议。
