乳及乳制品中阪崎肠杆菌PCR-DHPLC检测新技术的建立

Construction of Identification of Enterobacter sakazakii in Milk and Diary Product with PCR-DHPLC

摘要:

为了应用PCR结合变性高效液相色谱(DHPLC)技术建立食品中阪崎肠杆菌的快速检测方法, 根据阪崎肠杆菌16S-23S rRNA特异基因序列的特点设计特异性引物, PCR扩增的产物经DHPLC技术进行快速检测。以阪崎肠杆菌等59株参考菌株做特异性试验; 阪崎肠杆菌菌株稀释成不同梯度, 做灵敏度试验, 结果表明该方法具有很好的特异性, 方法灵敏度较高, 检测低限可达到为25 CFU/mL; 该方法可以快速、准确检测阪崎肠杆菌, 是食品中致病菌快速检测的新技术。

Abstract:

To identify the 16S-23S rRNA gene in Enterobacter sakazakiis, a PCR-DHPLC assay was performed in this study. Primers specific for the 16S-23S rRNA gene of Enterobacter sakazakii in food were selected to conduct the DHPLC assays. The specific testing was performed with Enterobacter sakazakii and 59 strains, and various grads of Enterobacter sakazakii was used for the sensitivity testing. The good specificity was found in the study. Also, the method showed nicer sensitivity with the lowest amount of detecting was 181 CFU/mL, so it can detect and identify Enterobacter sakazakii quickly and correctly. The DHPLC could be a new method for detecting food pathogens.