Anis Nagham, Bonifait Laetitia, Quesne Ségolène, Baugé Louise, Chemaly Marianne, Guyard-Nicodème Muriel
Unit for Hygiene and Quality of Poultry and Pork Products, Laboratory of Ploufragan-Plouzané-Niort, ANSES, 22440 Ploufragan, France.
Pathogens. 2023 Feb 16;12(2):338. doi: 10.3390/pathogens12020338.
In Europe, there is a process hygiene criterion for and on broiler carcasses after chilling. The criterion gives indicative contamination values above which corrective actions are required by food business operators. The reference methods for verifying compliance with the criterion for and are international standards EN ISO 6579-1 (2017) and EN ISO 10272-2 (2017), respectively. These methods are time-consuming and expensive for food business operators. Therefore, it would be advantageous to simultaneously detect spp. and quantify in the same analysis, using the same sample after the pre-enrichment step for recovery. A duplex PCR for detection and spp. enumeration was developed. Considering the method as a whole, the LOD and LOQ for enumeration were slightly over the limit of 3 log CFU/g set by the process hygiene criterion. A comparison of the duplex PCR method developed with the ISO method on artificially contaminated bacterial suspensions and on naturally contaminated samples demonstrated a good correlation of the results for enumeration when the duplex PCR was performed on samples taken before or after the pre-enrichment step, but revealed a slight bias with a large standard deviation resulting in widely spaced limits of agreement.
在欧洲，有一项关于冷却后肉鸡胴体上[具体细菌名称1]和[具体细菌名称2]的加工卫生标准。该标准给出了指示性污染值，超过此值食品企业经营者需采取纠正措施。用于验证是否符合[具体细菌名称1]和[具体细菌名称2]标准的参考方法分别是国际标准EN ISO 6579-1（2017）和EN ISO 10272-2（2017）。这些方法对食品企业经营者来说既耗时又昂贵。因此，在预富集步骤用于[具体细菌名称1]回收后，使用相同样本在同一次分析中同时检测[具体细菌名称1]并对[具体细菌名称2]进行定量将是有利的。开发了一种用于检测[具体细菌名称1]和计数[具体细菌名称2]的双重PCR方法。从整体方法来看，[具体细菌名称2]计数的检测限和定量限略高于加工卫生标准设定的3 log CFU/g的限值。将开发的双重PCR方法与ISO方法在人工污染的细菌悬液和天然污染的样本上进行比较，结果表明，当在预富集步骤之前或之后采集的样本上进行双重PCR时，[具体细菌名称2]计数结果具有良好的相关性，但显示出轻微偏差，标准偏差较大，导致一致性界限间隔较宽。
