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目的使用免疫磁分离-双重荧光定量PCR方法实现对鲜猪肉中金黄色葡萄球菌和志贺氏菌的同时、快速检测。方法在37℃的条件下,利用特异性免疫磁球从250 m L循环体系中快速、有效地捕获目标菌。再通过特异性的引物与探针,对沙门氏菌和志贺氏菌进行双重荧光定量PCR检测。结果本研究方法针对鲜猪肉中沙门氏菌和志贺氏菌的检测限分别达到3.4 cfu/g和9.4 cfu/g。方法总体灵敏度、特异性和准确度均达到100%。此外,通过对30组实际样品的检测,该方法与传统标准方法的结果保持一致。结论本研究建立的免疫磁分离-双重荧光定量PCR方法比国标方法更快速和高效,适用于鲜猪肉中沙门氏菌和志贺氏菌的快速检测。
Objective To detect the simultaneous and rapid detection of Staphylococcus aureus and Shigella in fresh pork by immunomagnetic separation - double fluorescence quantitative PCR. Methods Target bacteria were rapidly and efficiently captured from a 250 mL circulating system at 37 ° C using a specific immunomagnetic sphere. Then through specific primers and probes, Salmonella and Shigella were detected by double fluorescence quantitative PCR. Results The detection limits of Salmonella and Shigella in fresh pork were 3.4 cfu / g and 9.4 cfu / g, respectively. The overall sensitivity, specificity and accuracy of the method are all 100%. In addition, this method is consistent with the results of the traditional standard methods by testing 30 actual samples. Conclusion The immunomagnetic separation-double fluorescence quantitative PCR method established in this study is faster and more efficient than the national standard method and is suitable for the rapid detection of Salmonella and Shigella in fresh pork.