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目的建立一种空肠弯曲菌定量检测方法,为食品安全风险评估提供准确的数据支持。方法参照ISO/TS 10272—3和GB/T 4789.9—2008,基于MPN原理建立空肠弯曲菌定量检测方法,对定量加标的生鸡肉和鸡粪样品进行不同增菌培养基、不同培养环境和不同培养方式优化比对研究,用加标回收试验对建立的方法进行验证。结果增菌培养基的优化选择:Preston肉汤对加标菌量10 cfu/g的生鸡肉和鸡粪样品进行空肠弯曲菌检测的平均值分别为12.26和10.96 MPN/g,Bolton肉汤检测的平均值分别为2.38和2.32 MPN/g,二者比较差异有统计学意义(P<0.05);微需氧环境的优化选择:用三气培养箱法、产气袋法、抽气换气法和烛缸法对加标生鸡肉和鸡粪中的空肠弯曲菌检测的平均值分别为12.26和12.12 MPN/g、12.26和10.96 MPN/g、12.26和12.86 MPN/g、10.82和12.12 MPN/g,和三气培养箱法两两相比差异无统计学意义(P>0.05);培养方式:静止培养法对加标生鸡肉和鸡粪空肠弯曲菌的检测值分别为15.8和15.2 MPN/g,振荡培养法检测值分别为11.36和8.72 MPN/g,二者比较差异有统计学意义(P<0.05);用该法对低、中、高浓度加标生鸡肉样品的回收率分别为115.25%、111.5%和95.0%。结论此方法可以对高污染样品中空肠弯曲菌进行准确定量,特异度高,值得推广应用。
Objective To establish a method for the quantitative detection of Campylobacter jejuni and provide accurate data support for food safety risk assessment. Methods According to ISO / TS 10272-3 and GB / T 4789.9-2008, a quantitative detection method of Campylobacter jejuni was established based on the principle of MPN, and different enrichment culture media, culture conditions and culture conditions Methods to optimize the comparison study, with the spike recovery test to verify the established method. Results Optimization of the enrichment medium: Preston broth average values of Campylobacter jejuni for raw chicken and chicken manure samples with spiked 10 cfu / g bacteria were 12.26 and 10.96 MPN / g, respectively, The mean values were 2.38 and 2.32 MPN / g, respectively, with significant differences between the two groups (P <0.05). Optimum selection of micro-aerobic environment: Three-gas incubator, And Candlesticks for Campylobacter jejuni in spiked chicken and chicken manure were 12.26 and 12.12 MPN / g, 12.26 and 10.96 MPN / g, 12.26 and 12.86 MPN / g, 10.82 and 12.12 MPN / g, respectively (P> 0.05). The culture method: static culture method for the detection of Campylobacter jejuni in spiked chicken and chicken manure were 15.8 and 15.2 MPN / g , Shaking culture method detection values were 11.36 and 8.72 MPN / g, the difference was statistically significant (P <0.05); with the method of low, medium and high concentrations of spiked chicken samples were 115.25 %, 111.5% and 95.0%. Conclusion This method can accurately measure Campylobacter jejuni in highly contaminated samples with high specificity and is worth popularizing.