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

被引:29
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
相关论文
共 40 条
[1]  
Abramson Joseph H, 2004, Epidemiol Perspect Innov, V1, P6, DOI 10.1186/1742-5573-1-6
[2]  
Al Dahouk Sascha, 2003, Clin Lab, V49, P487
[3]  
Alton G, 1988, TECHNIQUES BRUCELLOS
[4]   Real-time PCR for identification of Brucella spp.: A comparative study of IS711, bcsp31 and per target genes [J].
Bounaadja, Lotfi ;
Albert, David ;
Chenais, Benoit ;
Henault, Sylvie ;
Zygmunt, Michel S. ;
Poliak, Sylvie ;
Garin-Bastuji, Bruno .
VETERINARY MICROBIOLOGY, 2009, 137 (1-2) :156-164
[5]   DIFFERENTIATION OF BRUCELLA-ABORTUS-BV-1, BRUCELLA-ABORTUS-BV-2, AND BRUCELLA-ABORTUS-BV-4, BRUCELLA-MELITENSIS, BRUCELLA-OVIS, AND BRUCELLA-SUIS-BV-1 BY PCR [J].
BRICKER, BJ ;
HALLING, SM .
JOURNAL OF CLINICAL MICROBIOLOGY, 1994, 32 (11) :2660-2666
[6]   Specificity of a polymerase chain reaction assay of a target sequence on the 31-kilodalton Brucella antigen DNA used to diagnose human brucellosis [J].
Casañas M.C. ;
Queipo-Ortuño M.I. ;
Rodriguez-Torres A. ;
Orduña A. ;
Colmenero J.D. ;
Morata P. .
European Journal of Clinical Microbiology and Infectious Diseases, 2001, 20 (2) :127-131
[7]   Detection of Brucella species DNA in the stomach content of aborted sheep fetuses by PCR [J].
Çetinkaya, B ;
Öngör, H ;
Muz, A ;
Ertas, HB ;
Kalender, H ;
Erdogan, HM .
VETERINARY RECORD, 1999, 144 (09) :239-240
[8]   DNA can interfere with detection of Borrelia burgdorferi in skin biopsy specimens by PCR [J].
Cogswell, FB ;
Bantar, CE ;
Hughes, TG ;
Gu, Y ;
Philipp, MT .
JOURNAL OF CLINICAL MICROBIOLOGY, 1996, 34 (04) :980-982
[9]  
Corbel M. J., 1985, Veterinary Bulletin, V55, P927
[10]   Detection of Brucella DNA from aborted bovine foetuses by polymerise chain reaction [J].
Cortez, A ;
Scarcelli, E ;
Soares, RM ;
Heinemann, MB ;
Sakamoto, SM ;
Genovez, ME ;
Perreira, P ;
Richtzenhain, LJ .
AUSTRALIAN VETERINARY JOURNAL, 2001, 79 (07) :500-501