title: Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples.
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-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.
authors.affiliation: Centro VISAVET, Universidad Complutense de Madrid, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Pérez-Sancho
authors.forename: T
authors.initials: T
authors.lastname: García-Seco
authors.forename: L
authors.initials: L
authors.lastname: Arrogante
authors.forename: N
authors.initials: N
authors.lastname: García
authors.forename: I
authors.initials: I
authors.lastname: Martínez
authors.forename: A
authors.initials: A
authors.lastname: Diez-Guerrier
authors.forename: A
authors.initials: A
authors.lastname: Perales
authors.forename: J
authors.initials: J
authors.lastname: Goyache
authors.forename: L
authors.initials: L
authors.lastname: Domínguez
authors.forename: J
authors.initials: J
authors.lastname: Alvarez
mesh_terms: D037881:Aborted Fetus / Q000382:microbiology; D000034:Abortion, Veterinary / Q000382:microbiology; D000818:Animals; D002002:Brucella / Q000302:isolation & purification*; D002006:Brucellosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary*; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial
keywords: Bacteriology; Brucella; Loop mediated isothermal amplification (LAMP); PCR