Comparison of PCR assay and bacteriological culture method for the detection of Brucella melitensis in stomach content samples of aborted sheep fetuses


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Ilhan Z., Solmaz H., Aksakal A., Gülhan T., Ekin İ. H., Boynukara B.

Deutsche Tierarztliche Wochenschrift, vol.114, no.12, pp.460-464, 2007 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 114 Issue: 12
  • Publication Date: 2007
  • Doi Number: 10.2377/0341-6593-114-460
  • Journal Name: Deutsche Tierarztliche Wochenschrift
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.460-464
  • Keywords: Bacteriological culture, Brucella melitensis, PCR, RBPT, Sheep
  • Ondokuz Mayıs University Affiliated: No

Abstract

The aim of this study was to evaluate the polymerase chain reaction (PCR) assay for detection of Brucella melitensis in stomach content samples of aborted sheep fetuses and to compare its performance with bacteriological culture method. It was also aimed to determine the agreement between PCR and Rose Bengal plate test (RBPT). Materials were collected from aborted sheep from 109 different sheep flocks in the region of Van during the lambing seasons of 2004-2005 and 2005-2006. Stomach contents from 135 aborted sheep fetuses were examined by bacteriological culture and PCR, and 135 sera from these aborting ewes were tested by RBPT Identification and typing of Brucella strains were performed using standard classification test. B. melitensis biovar 3 was isolated from 26 (19.2 %) of foetal stomach contents. B. melitensis was detected by PCR in 29 (21.4 %) stomach content samples. Twenty five sera (18.5 %) from aborting ewes tested positive by RBPT. The detection limit of B. melitensis 16 M strain by PCR was 1.7 × 103 cfu (colony forming units)/ml in spiked stomach contents. Diagnostic sensitivity and specificity of the PCR were detected as 100 % and 97.2 %, respectively. The agreement between PCR and RBPT was found to be 97 %. In conclusion, PCR assay would have an advantage over conventional bacteriological culture method, but in particular for its ability to meet the specificity reguirements for the detection of B. melitensis in stomach content samples of aborted sheep fetuses. © 2007 M. & H. Schaper GmbH.