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为实现奶粉中常见污染菌穆汀斯克罗诺杆菌的快速检测与控制,本文根据cgcA基因序列设计出特异性探针和引物,建立了运用实时荧光定量PCR法检测穆汀斯克罗诺杆菌的反应体系和反应条件,并对该法的特异性、灵敏度、稳定性进行评价,并进行人工染菌样品检测实验。特异性结果表明,对穆汀斯克罗诺杆菌的荧光PCR检测结果的特异性为100%;灵敏度结果表明,穆汀斯克罗诺杆菌检出限在440 cfu/m L;稳定性结果表明,穆汀斯克罗诺杆菌组内实验CV在0.48~0.69%之间波动,而组间实验CV在0.69~0.73%之间波动;人工染菌样品试验表明,在增菌6 h后能检出阳性。本研究所建立的实时荧光定量PCR法特异性好、灵敏度高、稳定性强,有望成为快速检测食品中穆汀斯克罗诺杆菌污染的新方法,具有很好的研究价值和应用前景。
In order to realize the rapid detection and control of Corynebacterium mucronella in milk powder, specific probes and primers were designed according to the cgcA gene sequence, and the real-time fluorescence quantitative PCR System and reaction conditions, and the specificity, sensitivity and stability of the method were evaluated, and the experiment of artificial dye-contaminated samples was carried out. The specificity of the results showed that the specificity of the PCR detection of Corynebacterium glutinosa was 100%. The sensitivity showed that the detection limit of Corynebacterium glutamate was 440 cfu / mL. The stability results showed that the sensitivity The CV of test group was fluctuated between 0.48 and 0.69%, while CV between the two groups fluctuated between 0.69 and 0.73%. The test of artificial bacterial strain showed that the CV of positive strain could be detected 6 hours after enrichment. The real-time fluorescence quantitative PCR method established in this study has good specificity, high sensitivity and strong stability. It is expected to become a new method to rapidly detect the contamination of Cromonus Rodinarius in food, which has good research value and application prospect.