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-IS. 711 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 IS. 711-LAMP technique for direct Brucella detection.
Bibliographical noteFunding Information:
This study was supported by AGL-2009-12674 project ( Ministerio de Ciencia e Innovación ). Marta Pérez-Sancho is a recipient of a predoctoral grant from Consejería de Educación de la Comunidad de Madrid (IV Plan regional de investigación científica e innovación tecnológica, 2005-2008) y Fondo Social Europeo. Authors also acknowledge the support of the EU FP7 ANTIGONE Project Number 278976 .
- Loop mediated isothermal amplification (LAMP)