乳粉中普通变形杆菌和奇异变形杆菌复合PCR-DHPLC检测技术的建立

Identification of Proteus vulgaris and Proteus Mirabilis in milk Powders by Multiplex PCR-DHPLC

应用复合PCR结合变性高效液相色谱(DHPLC)技术,建立乳粉中普通变形杆菌和奇异变形杆菌的高通量检测方法。根据普通变形杆菌的blaA和blaB基因及奇异变形杆菌的ureR基因序列分别设计特异性引物,复合PCR扩增的产物经DHPLC技术进行快速检测,并以37株参考菌株做特异性试验。试验结果表明,该方法具有很好的特异性,经复合PCR-DHPLC可同时检测乳粉中普通变形杆菌和奇异变形杆菌。该方法可以快速、准确、高通量检测普通变形杆菌和奇异变形杆菌,是乳粉中致病菌高通量检测的新技术。

To identify Proteus vulgaris and Proteus mirabili in milk powders,a multiplex PCR-Denaturing High-Performance Liquid Chromatography(DHPLC)assay was performed in this study.Primers specific for blaA and blaB gene of Proteus vulgaris and ureR gene of Proteus mirabilis were selected to conduct the multiplex PCR-DHPLC assays.The specific testing was performed with 37 strains.The good specificity was found in the study.Also,it can detect and identify Proteus vulgaris and Proteus mirabilis quickly and highthrough by a multiplex PCR-DHPLC.The DHPLC could be a new method for detecting pathogens in milk powders.