4.6 Article

Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples

Journal

RESEARCH IN VETERINARY SCIENCE
Volume 95, Issue 2, Pages 489-494

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/j.rvsc.2013.05.002

Keywords

Bacteriology; Brucella; Loop mediated isothermal amplification (LAMP); PCR

Funding

  1. Ministerio de Ciencia e Innovacion [AGL-2009-12674]
  2. Consejeria de Educacion de la Comunidad de Madrid (IV Plan regional de investigacion cientifica e innovacion tecnologica)
  3. Fondo Social Europeo
  4. EU FP7 ANTIGONE Project [278976]

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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. (c) 2013 Elsevier Ltd. All rights reserved.

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