{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T03:29:27Z","timestamp":1789615767756,"version":"build-2803163510"},"reference-count":22,"publisher":"Canadian Science Publishing","issue":"5","license":[{"start":{"date-parts":[[2008,5,1]],"date-time":"2008-05-01T00:00:00Z","timestamp":1209600000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/www.nrcresearchpress.com\/page\/about\/CorporateTextAndDataMining"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Can. J. Microbiol."],"published-print":{"date-parts":[[2008,5]]},"abstract":"<jats:p>Brucellosis is a widespread zoonosis. Currently the diagnosis of this zoonosis is based on microbiological and serological laboratory tests. Polymerase chain reaction (PCR) has been used to detect DNA from Brucella . Different target genes, primer pairs, PCR techniques, and extraction procedures have previously been published for Brucella detection. But only a few of these primers have been used in human samples, and only one study has been carried out to compare sensitivity between them. In the present study, 3 sets of primers and 3 different PCR protocols amplifying 3 different regions of the Brucella genome were compared for detection of Brucella DNA in a peripheral-blood PCR assay to conclude which is most suitable for the clinical diagnostic laboratory. These 3 pairs of primers amplify 3 different fragments included in (i) a gene encoding a 31\u00a0kDa Brucella abortus antigen (B4\/B5), (ii) a sequence 16S rRNA of B.\u00a0abortus (F4\/R2), and (iii) a gene encoding an outer membrane protein (omp-2) (JPF\/JPR). Some modifications on the reported techniques were applied during the present work to improve the outcome. The results showed that the B4\/B5 primer pair had the highest sensitivity for detection of positive samples (98%), the JPF\/JPR primer pair detected 88.4% of positive samples, whereas F4\/R2 primer pair was the least sensitive, being able to detect only 53.1% of positive samples. The specificity of the 3 techniques was 100%. The B4\/B5 primer pair was also able to detect the smallest number of bacteria (700 cfu\/mL), whereas JPF\/JPR was able to detect 7\u00a0\u00d7 10<jats:sup>5<\/jats:sup>\u00a0cfu\/mL and F4\/R2 was able to detect 7\u00a0\u00d7 10<jats:sup>7<\/jats:sup>cfu\/mL. It is thus concluded that using the B4\/B5 primer PCR with the suggested modifications is a robust assay, which meets the sensitivity requirements to be used for testing of human blood samples for brucellosis in the diagnostic laboratory.<\/jats:p>","DOI":"10.1139\/w08-017","type":"journal-article","created":{"date-parts":[[2008,4,30]],"date-time":"2008-04-30T18:11:49Z","timestamp":1209579109000},"page":"352-357","source":"Crossref","is-referenced-by-count":36,"title":["Evaluation of three polymerase chain reaction techniques for detection of<i>Brucella<\/i>DNA in peripheral human blood"],"prefix":"10.1139","volume":"54","author":[{"given":"Manal M.","family":"Baddour","sequence":"first","affiliation":[{"name":"Microbiology and Immunology Department, Faculty of Medicine, Alexandria University."},{"name":"Girls College of Education, Riyadh College, Riyadh, Saudi Arabia."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dalal H.","family":"Alkhalifa","sequence":"additional","affiliation":[{"name":"Microbiology and Immunology Department, Faculty of Medicine, Alexandria University."},{"name":"Girls College of Education, Riyadh College, Riyadh, Saudi Arabia."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"155","reference":[{"key":"atypb1\/ref1","first-page":"487","volume":"49","author":"Al Dahouk S.","year":"2003","journal-title":"Clin. 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