Centro VISAVET, Universidad Complutense de Madrid, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain.
Res Vet Sci. 2013 Oct;95(2):489-94. doi: 10.1016/j.rvsc.2013.05.002. Epub 2013 May 25.
DNA-based methods have emerged as an additional tool for Brucella infection-confirmation at a herd level. However, their implementation may require the use of specialized equipment. In this context the recently developed loop-mediated isothermal amplification (LAMP) technique may constitute an additional and cost-effective tool for rapid and specific DNA detection, especially in low income areas. In the present study the usefulness of a newly developed LAMP assay aiming at the multicopy-IS711 sequence was assessed on a variety of clinical samples (n=81 from abortions and ewes; cattle, n=3; swine, n=4) that were analyzed in parallel using real-time PCR and bacteriology. Although overall sensitivities obtained with the three methods were comparable (p>0.05), our results highlighted the complementarity between bacteriology and molecular-based methods for increased sensitivity. Significant differences (p<0.05) were observed with all techniques depending on the nature of the sample. Our results demonstrate the potential of the IS711-LAMP technique for direct Brucella detection.
基于 DNA 的方法已成为在畜群水平上确认布鲁氏菌感染的另一种工具。然而，它们的实施可能需要使用专门的设备。在这种情况下，最近开发的环介导等温扩增 (LAMP) 技术可能是一种额外的、具有成本效益的快速和特异性 DNA 检测工具，特别是在低收入地区。在本研究中，评估了一种新开发的针对多拷贝 IS711 序列的 LAMP 检测方法在各种临床样本（流产和母羊 n=81；牛 n=3；猪 n=4）中的应用，这些样本同时使用实时 PCR 和细菌学进行分析。尽管三种方法的总体灵敏度相当（p>0.05），但我们的结果强调了细菌学和基于分子的方法之间的互补性，以提高灵敏度。根据样本的性质，所有技术都观察到显著差异（p<0.05）。我们的结果证明了 IS711-LAMP 技术用于直接布鲁氏菌检测的潜力。
