3种方法鉴定鸡肉粉中沙门氏菌的比对研究

Comparative study on the identification of Salmonella in chicken powder by 3 methods

中文关键词:  沙门氏菌  实时荧光定量PCR法  基质辅助激光解吸电离飞行时间质谱法  血清分型  鉴定

英文关键词:Salmonella  quantitative real-time PCR  matrix-assisted laser desorption ionization time-of-flight mass spectrometry  serotyping  identification

中文摘要:

      目的 采用3种方法对鸡肉粉中沙门氏菌进行检测。方法 3份样品的前处理依据组织方提供的《作业指导书》进行, 每瓶样品直接加入4.5 mL灭菌去离子水复溶, 作为初始样本, 后续实验依据GB 4789.4—2016《食品安全国家标准 食品微生物学检验 沙门氏菌检验》操作。鉴定分离出疑似菌后, 加入了实时荧光定量PCR法、基质辅助激光解吸电离飞行时间质谱法2种方法作为辅助检测, 并用全自动微生物鉴定系统进行生化鉴定, 再结合生化鉴定与血清学实验综合评判检测结果。结果 使用3种鉴定方法在JS015、JS110、JS140样本中均检出沙门氏菌。结论 本研究采用的实时荧光定量PCR法、基质辅助激光解吸电离飞行时间质谱法2种方法均能快速准确鉴定出沙门菌, 特别是基质辅助激光解吸电离飞行时间质谱法和血清学分型配合使用, 对沙门菌的快速鉴定有重要意义。

英文摘要:

      Objective To compare the identification of Salmonella in chicken powder by 3 methods. Methods The pretreatment of the 3 samples was conducted according to the Work instruction provided by the organizer. Each bottle of samples was directly added into 4.5 mL sterilized deionized water for re-dissolution as the initial samples, and the subsequent experiments were conducted in accordance with GB 4789.4—2016 National food safety standard-Food microbiological examination-Salmonella. After identification of the suspected bacteria, quantitative real-time PCR and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) were added as the auxiliary detection methods. Biochemical identification was carried out with the automatic microbial identification system, and the detection results were comprehensively evaluated with the combination of biochemical identification and serological experiment. Results Salmonella was detected in samples JS015, JS110 and JS140 by 3 identification methods. Conclusion The quantitative real-time PCR method and MALDI-TOF-MS method are able to identify Salmonella quickly and accurately. In particular, MALDI-TOF-MS method and serum credit type are used in combination, which is of great significance for the rapid identification of Salmonella.