一种简化的方法，用于检测屠宰猪扁桃体中的致病性肠侵袭性大肠杆菌。
A simplified method for detecting pathogenic Yersinia enterocolitica in slaughtered pig tonsils.
机构信息
Anses (Agence nationale de sécurité sanitaire), unité HQPAP, BP53, 22440 Ploufragan, France.
出版信息
J Microbiol Methods. 2010 Nov;83(2):244-9. doi: 10.1016/j.mimet.2010.09.012. Epub 2010 Sep 17.
The aim of this study was to collect preliminary data on the carriage of pathogenic Yersinia enterocolitica in slaughtered pigs in France and to test a simplified method for detecting these strains from tonsils. From January to March 2009, 900 tonsil swabs were taken from pigs at one slaughterhouse in Brittany, France. The swabs were vortexed in 10 ml PSB broth, then 1 ml was added to 9 ml ITC broth. The media were incubated for 48 h at 25°C. The PSB enrichment broth was streaked on CIN plates and the ITC enrichment broth on SSDC plates. In addition to the ISO 10273 method, we also streaked ITC enrichment broth on CIN plates. The plates were incubated for 24h at 30°C, and we then streaked a maximum of four typical colonies per plate onto a plate containing chromogenic medium (YeCM), for the isolation of pathogenic Y. enterocolitica isolates. In parallel, biochemical assays were carried out to confirm the identification of the isolates as Yersinia and to determine biotype. After passage on a YeCM plate and biochemical tests, 380 strains were confirmed to be pathogenic Y. enterocolitica. Finally, with the ISO 10273 method, 9.1% (CI(95%) [5.8-12.4]) of tonsil swabs and 60% (CI(95%) [45.4-74.6]) of the batches were positive. With the ITC-CIN method, 14.0% (CI95% [10.7-17.3]) of the tonsil swabs and 68.9% (CI(95%) [54.3-83.5]) of the batches were positive. Identification as pathogenic Y. enterocolitica was confirmed for 97.0% of the typical colonies obtained on the chromogenic medium, YeCM. The most prevalent biotype was biotype 4 (80.5% of the isolates), followed by biotype 3. This study demonstrates that the ITC-CIN method, followed by streaking on YeCM, may be an effective approach to the isolation of pathogenic Y. enterocolitica from tonsil swabs and the recovery of positive samples. This method is less time-consuming than the ISO 10273 method and reduces the number of biochemical tests required for the confirmation of Yersinia identification, through the use of YeCM.
本研究旨在收集法国屠宰场猪源致病性小肠结肠炎耶尔森菌带菌情况的初步数据，并对一种从扁桃体中检测这些菌株的简化方法进行测试。2009 年 1 月至 3 月，从法国布列塔尼的一个屠宰场采集了 900 个扁桃体拭子。将拭子在 10mlPSB 肉汤中涡旋，然后向 9mlITC 肉汤中加入 1ml。将培养基在 25°C 下孵育 48 小时。PSB 富集肉汤在 CIN 平板上划线，ITC 富集肉汤在 SSDC 平板上划线。除了 ISO10273 方法外，我们还在 CIN 平板上划线 ITC 富集肉汤。平板在 30°C 下孵育 24 小时，然后从每个平板上最多划线 4 个典型菌落到含有显色培养基（YeCM）的平板上，用于分离致病性小肠结肠炎耶尔森菌。同时，进行生化检测以确认分离物为耶尔森菌，并确定生物型。在 YeCM 平板上传代和生化试验后，380 株被确认为致病性小肠结肠炎耶尔森菌。最后，采用 ISO10273 方法，9.1%（95%CI [5.8-12.4]）的扁桃体拭子和 60%（95%CI [45.4-74.6]）的批次为阳性。采用 ITC-CIN 方法，14.0%（95%CI [10.7-17.3]）的扁桃体拭子和 68.9%（95%CI [54.3-83.5]）的批次为阳性。在显色培养基 YeCM 上获得的典型菌落中，97.0%被确认为致病性小肠结肠炎耶尔森菌。最常见的生物型是 4 型（80.5%的分离株），其次是 3 型。本研究表明，ITC-CIN 方法，继之以在 YeCM 平板上划线，可能是从扁桃体拭子中分离致病性小肠结肠炎耶尔森菌和回收阳性样本的有效方法。该方法比 ISO10273 方法耗时更少，并且通过使用 YeCM，减少了确认耶尔森菌鉴定所需的生化检测次数。
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