RT-PCR快速检测婴幼儿奶粉中的克罗诺杆菌活菌研究

RT-PCR快速检测婴幼儿奶粉中的克罗诺杆菌活菌研究
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。

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.

标题：RT-PCR快速检测婴幼儿奶粉中的克罗诺杆菌活菌研究
英文标题：Rapid detection of Cronobacter in infant milk powder by reverse transcription PCR

作者：
孙晶莹 陕西省人民医院
李宏铎 西安市食品药品检验所
孙丽君 陕西省人民医院
杨晓东 西安市食品药品检验所
胡军 陕西省人民医院

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,

发表日期：2017-10-16

2025-1-31 19:53 上传 RT-PCR快速检测婴幼儿奶粉中的克罗诺杆菌活菌研究.pdf

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