• 安全检测 •

### 3 种致病菌多重real-time PCR检测方法的建立及其在散装即食肉制品中的应用

1. （中国肉类食品综合研究中心，北京食品科学研究院，北京 100068）
• 出版日期:2022-01-25 发布日期:2022-01-29
• 基金资助:
“十三五”国家重点研发计划重点专项（2018YFC1603400）； 北京市优秀人才培养资助青年骨干个人项目（2017754154700G101）

### Establishment and Application of Multiple Real-time PCR Method for Detection of Three Kinds of Pathogenic Bacteria in Bulk Ready-to-eat Meat Products

FAN Wei, GAO Xiaoyue, LI Henan, DONG Yuxin, LI Yuxuan, LIU Hongyu, GUO Wenping

1. (China Meat Research Center, Beijing Academy of Food Sciences, Beijing 100068, China)
• Online:2022-01-25 Published:2022-01-29

Abstract: A triplex real-time polymerase chain reaction (real-time PCR) method for rapid simultaneous detection of Salmonella (SAL), Staphylococcus aureus (SA) and Bacillus cereus (BC) was established. Primers and TaqMan probes were designed targeting the invA gene of SAL, the Sa442 gene of SA, and the Cereolysin AB gene of BC. The performance indicators of the developed method were evaluated, and actual samples of ready-to-eat meat products were detected by it. The results showed that the real-time PCR method was highly reproducible, specific and sensitive. Fifteen non-target reference strains were tested by this method and the results were all negative. The standard curves for the three pathogenic bacteria were linear over the concentration range of 102–108 CFU/mL, and the intra-assay and inter-assay coefficients of variation (CVs) were less than 2% in the quantitative tests. Without bacterial enrichment, the limits of detection (LOD) for SAL, SA, and BC were 3.8 × 102, 4.9 × 102, and 5.7 × 102 CFU/mL, respectively. After 5 h enrichment, they increased to 3.8, 4.9, and 5.7 CFU/mL, respectively. The results of the real-time PCR method for 100 actual samples were consistent with those of the national standard method, indicating that the real-time PCR method can be used for the detection of the three pathogenic bacteria in bulk ready-to-eat meat products.