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

被引:27
|
作者
Perez-Sancho, M. [1 ,2 ]
Garcia-Seco, T. [1 ]
Arrogante, L. [3 ]
Garcia, N. [1 ]
Martinez, I. [1 ]
Diez-Guerrier, A. [2 ]
Perales, A. [3 ]
Goyache, J. [1 ,2 ]
Dominguez, L. [1 ,2 ]
Alvarez, J. [1 ,4 ]
机构
[1] Univ Complutense Madrid, Ctr VISAVET, E-28040 Madrid, Spain
[2] Univ Complutense Madrid, Fac Vet, Dept Sanidad Anim, E-28040 Madrid, Spain
[3] Minist Agr Alimentac & Media Ambiente, Lab Cent Sanidad Anim, Granada 18320, Spain
[4] IRYCIS, Madrid 28034, Spain
关键词
Bacteriology; Brucella; Loop mediated isothermal amplification (LAMP); PCR; POLYMERASE-CHAIN-REACTION; REAL-TIME PCR; ABORTED BOVINE FETUSES; RAPID DETECTION; REEMERGING ZOONOSIS; DNA; MILK; MELITENSIS; BLOOD; SHEEP;
D O I
10.1016/j.rvsc.2013.05.002
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
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.
引用
收藏
页码:489 / 494
页数:6
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