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首页> 外文期刊>Animal Biotechnology >Loop-Mediated Isothermal Amplification for Rapid Detection and Differentiation of Wild-Type Bovine Herpesvirus-1 and Glycoprotein E-Deleted Marker Vaccine Strain
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Loop-Mediated Isothermal Amplification for Rapid Detection and Differentiation of Wild-Type Bovine Herpesvirus-1 and Glycoprotein E-Deleted Marker Vaccine Strain

机译：循环介导的等温扩增，用于快速检测和区分野生型牛疱疹病毒1和糖蛋白E缺失的标记疫苗株

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Bovine herpesvirus-1 (BoHV-1) is an important viral pathogen affecting cattle and causing numerous reproductive disorders leading to significant economic losses to the cattle industry. The control programs for BoHV-1 are widely based on the use of glycoprotein E-deleted marker vaccines, wherein detection and differentiation of wild-type and gE-deleted vaccine strains is of crucial importance for proper disease management. In this study, we report rapid and simple loop-mediated isothermal amplification (LAMP) assays for detection and differentiation of gE-deleted BoHV-1 from wild-type virus under isothermal conditions. The assays could be completed in 90 mintes, including viral DNA isolation, target amplification and visual interpretation of results with naked eye. The analytical sensitivity of the assays was 10 times higher than conventional PCR and could detect as little as 100 fg of viral DNA per reaction. The applicability of LAMP for detection of BoHV-1 in bovine semen was assessed by testing semen samples collected from breeding bulls and compared with TaqMan real-time PCR (as gold standard). The LAMP assays had diagnostic specificity of 100%. The diagnostic sensitivity was 88.2% and 83.3% for gB-and gE-LAMP, respectively, when compared with TaqMan real-time PCR. Our results have shown that the LAMP method developed in this study is a potential tool for rapid, sensitive, specific, cost-effective, and user-friendly detection and differentiation of wild type BoHV-1 from gE-deleted marker vaccine.
机译：牛疱疹病毒1（BoHV-1）是一种重要的病毒病原体，影响牛，并引起许多繁殖障碍，给牛业造成重大经济损失。 BoHV-1的控制程序广泛基于缺失糖蛋白E的标记疫苗的使用，其中检测和区分野生型和gE缺失的疫苗株对于正确的疾病管理至关重要。在这项研究中，我们报告了快速和简单的回路介导的等温扩增（LAMP）分析方法，用于在等温条件下从野生型病毒中检测和区分gE缺失的BoHV-1。该检测可在90分钟内完成，包括病毒DNA分离，靶标扩增和肉眼可视化结果分析。该方法的分析灵敏度比常规PCR高10倍，每个反应可检测到100 fg的病毒DNA。通过测试从种公牛收集的精液样本并与TaqMan实时PCR（作为金标准）进行比较，评估了LAMP在牛精液中检测BoHV-1的适用性。 LAMP分析的诊断特异性为100％。与TaqMan实时PCR相比，gB-和gE-LAMP的诊断敏感性分别为88.2％和83.3％。我们的结果表明，在这项研究中开发的LAMP方法是一种快速，灵敏，特异，具有成本效益且用户友好的方法，可从gE缺失标记疫苗中检测和区分野生型BoHV-1，这是一种潜在的工具。

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