Ongör H, Cetinkaya B, Karahan M, Bulut H
Department of Microbiology, Faculty of Veterinary Medicine, University of Firat, Elazig, Turkey.
Foodborne Pathog Dis. 2006 Fall;3(3):245-50. doi: 10.1089/fpd.2006.3.245.
The current study was carried out to assess the use of immunomagnetic separation-polymerase chain reaction (IMS-PCR) in direct detection of Brucella abortus and B. melitensis from soft cheese and to examine a relatively small number of field samples for the presence of these species. Two methodologies, one with IMS and the other without IMS, were employed for recovery of the Brucella species from cheese samples. IMS in conjunction with the PCR assay was determined to detect as low as 3x10(2) bacteria/mL, while the limit of detection with the other extraction procedure was 3x10(3) bacteria/mL. In the analysis of 40 cheese samples collected from various markets, only B. abortus was detected by PCR using both DNA extraction procedures in two (5%) samples. No positive results were obtained by culture and B. melitensis was not found in any cheese samples examined. The results suggest that this technique is promising owing to its pace and high sensitivity and should aid in direct detection of Brucella species from complex food samples.
本研究旨在评估免疫磁珠分离-聚合酶链反应（IMS-PCR）用于直接检测软质奶酪中流产布鲁氏菌和羊种布鲁氏菌的效果，并检测相对少量的现场样本中这些菌种的存在情况。采用两种方法从奶酪样本中回收布鲁氏菌，一种采用免疫磁珠分离法，另一种不采用。免疫磁珠分离法结合PCR检测法可检测低至3×10² 个细菌/毫升，而另一种提取方法的检测限为3×10³ 个细菌/毫升。在分析从不同市场收集的40份奶酪样本时，使用两种DNA提取方法通过PCR检测，仅在两份（5%）样本中检测到流产布鲁氏菌。培养未获得阳性结果，在所检测的任何奶酪样本中均未发现羊种布鲁氏菌。结果表明，该技术由于其速度和高灵敏度而具有前景，应有助于从复杂食品样本中直接检测布鲁氏菌。
