caa a22 4500 463238524 CHVBK 20180405153305.0 cr unu---uuuuu 170326e20071101xx s 000 0 eng 10.1007/s11259-006-0109-6 doi (NATIONALLICENCE)springer-10.1007/s11259-006-0109-6 Diagnosis of Canine Brucellosis: Comparison between Serological and Microbiological Tests and a PCR Based on Primers to 16S-23S rDNA Interspacer [Elektronische Daten] [L. Keid, R. Soares, N. Vieira, J. Megid, V. Salgado, S. Vasconcellos, M. da Costa, F. Gregori, L. Richtzenhain] A pair of primers directed to 16S-23S rDNA interspacer (ITS) was designed directed to Brucella genetic sequences in order to develop a polymerase chain reaction (PCR) putatively capable of amplifying DNA from any Brucella species. Nucleic acid extracts from whole-blood from naive dogs were spiked with decreasing amounts of Brucella canis RM6/66 DNA and the resulting solutions were tested by PCR. In addition, the ability of PCR to amplify Brucella spp. genetic sequences from naturally infected dogs was evaluated using 210 whole-blood samples of dogs from 19 kennels. The whole-blood samples collected were subjected to blood culture and PCR. Serodiagnosis was performed using the rapid slide agglutination test with and without 2-mercaptoethanol. The DNA from whole blood was extracted using proteinase-K, sodium dodecyl sulphate and cetyl trimethyl ammonium bromide followed by phenol-chloroform purification. The PCR was capable of detecting as little as 3.8 fg of Brucella DNA mixed with 450 ng of host DNA. Theoretically, 3.8 fg of Brucella DNA represents the total genomic mass of fewer than two bacterial cells. The PCR diagnostic sensitivity and specificity were 100%. From the results observed in the present study, we conclude that PCR could be used as confirmatory test for diagnosis of B. canis infection. Springer Science + Business Media, Inc., 2007 Brucella canis nationallicence canine brucellosis nationallicence PCR nationallicence internal transcribed spacer nationallicence blood culture nationallicence rapid slide agglutination test nationallicence CTAB : cetyl trimethyl ammonium bromide nationallicence ITS : interspacer nationallicence PCR : polymerase chain reaction nationallicence RBPT : Rose Bengal Plate test nationallicence R-LPS : rough lipopolysaccharide nationallicence RSAT : rapid slide agglutination test nationallicence SDS : sodium docdecyl sulphate nationallicence Keid L. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut Soares R. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut Vieira N. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut Megid J. Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine, São Paulo State University, Botucatu, São Paulo, Brazil aut Salgado V. Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine, São Paulo State University, Botucatu, São Paulo, Brazil aut Vasconcellos S. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut da Costa M. Departament of Microbiology, Health Sciences Institute, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil aut Gregori F. Research and Developmental Center of Animal Health, Biological Institute, São Paulo, São Paulo, Brazil aut Richtzenhain L. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut Veterinary Research Communications Springer Netherlands 31/8(2007-11-01), 951-965 0165-7380 31:8<951 2007 31 11259 https://doi.org/10.1007/s11259-006-0109-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11259-006-0109-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Keid L. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut NATIONALLICENCE 700 1- Soares R. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut NATIONALLICENCE 700 1- Vieira N. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut NATIONALLICENCE 700 1- Megid J. Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine, São Paulo State University, Botucatu, São Paulo, Brazil aut NATIONALLICENCE 700 1- Salgado V. Department of Veterinary Hygiene and Public Health, School of Veterinary Medicine, São Paulo State University, Botucatu, São Paulo, Brazil aut NATIONALLICENCE 700 1- Vasconcellos S. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut NATIONALLICENCE 700 1- da Costa M. Departament of Microbiology, Health Sciences Institute, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil aut NATIONALLICENCE 700 1- Gregori F. Research and Developmental Center of Animal Health, Biological Institute, São Paulo, São Paulo, Brazil aut NATIONALLICENCE 700 1- Richtzenhain L. Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine, University of São Paulo, São Paulo, SP, Brazil aut NATIONALLICENCE 773 0- Veterinary Research Communications Springer Netherlands 31/8(2007-11-01), 951-965 0165-7380 31:8<951 2007 31 11259 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer