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

M. Pérez-Sancho, T. García-Seco, L. Arrogante, N. García, I. Martínez, A. Diez-Guerrier, A. Perales, J. Goyache, L. Domínguez, J. Álvarez

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)489-494
Number of pages6
JournalResearch in veterinary science
Volume95
Issue number2
DOIs
StatePublished - Oct 2013

Bibliographical note

Funding 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 .

Keywords

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

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