RT-PCR快速检测婴幼儿奶粉中的克罗诺杆菌活菌研究

Rapid detection of Cronobacter in infant milk powder by reverse transcription PCR

中文关键词:  反转录PCR  克罗诺杆菌  活菌  奶粉

英文关键词:reverse transcription PCR  Cronobacter  live bacteria  milk powder

中文摘要:

      目的 建立反转录PCR(reverse transcription PCR, RT-PCR)快速检测克罗诺杆菌的方法, 解决常见方法中以DNA作为模板带来的假阳性问题。方法 根据克罗诺杆菌的α-葡萄糖苷酶基因序列设计特异性引物和探针, 检测克罗诺杆菌, 做了特异性和灵敏度研究。结果 建立的RT-PCR法能有效扩增克罗诺杆菌的特异性片段, 在纯培养物中, 灵敏度可达10 CFU/mL, 在人工污染奶粉培养7 h后的检测限为1×10 CFU/mL。 结论 本方法能够快速检测奶粉中克罗诺杆菌活菌。

英文摘要:

      Objective To establish a method for rapid detection of viable Cronobacter by reverse transcription PCR (RT-PCR), in order to solve the false positive problem with DNA as template in common methods. Methods According to α-glucosidase gene sequence of the Cronobacter, specific primers and probes was designed for the determination of Cronobacter. The sensitivity and specificity of the RT-PCR method were compared with conventional methods. Results The RT-PCR method could effectively amplified specific fragment of Cronobacter. In the pure culture, the sensitivity was up to 10 CFU/mL. The limit of detection of the artificial milk powder after 7 h was 1×10 CFU/mL. Conclusion This method can rapidly detect the viable Cronobacter in the powdered milk.