摘要
目的:由于中国药典中规定的沙门菌检查采用微生物培养法,其操作繁琐、培养周期长,本研究拟建立一种快速定性检测沙门菌的方法以替代药典中繁琐耗时的微生物培养法。方法:取10 m L含动物类药材的口服制剂,分别加入0.096~96 cfu的沙门菌,同时以大肠埃希菌作为干扰对照菌,设置沙门菌污染组、大肠埃希菌污染组、沙门菌及大肠埃希菌混合污染组及阴性对照组共4个实验组,采用多重聚合酶链扩增技术(PCR)对供试品溶液进行扩增检测,分别考察该方法对沙门菌检出的专属性、准确性、灵敏度以及适用性。结果:所建立的方法检验周期短,仅需30小时;专属性好,能准确区分沙门菌与干扰对照菌;结果准确,检测结果与药典方法检验结果一致;灵敏度高,最低检测限为1 cfu。结论:本方法便捷高效、结果准确,可为药品检验中的沙门菌检查提供一种新手段。
Objective: Microbial culture method is tedious and time-consuming, which is prescribed in the Chinese Pharmacopoeia for Salmonella detection, this study intends to establish a rapid qualitative method to replace microbial culture method. Methods: Adding 0.096-96 cfu of Salmonella into 10 mL oral medicines containing animal ingredients, and E. coli as interference bacteria. Four experimental groups were setted i.e.E, coli contamination group, Salmonella contamination group, E. coil and Salmonella contamination group and negative control group, Salmonella possible in drugs was detected by multiplex polymerase chain reaction ( PCR ). Some experimental factors were investaged such as the specificity, accuracy, sensitivity and applicability. Results: Salmonella can be detected as low as 1 cfu within 30 hours even in the presence of other bacteria, and the results were consistent with those of the pharmacopoeia method. Conclusions: The method may be an alternative for Salmonella detection for its high efficiency and accuracy.
出处
《现代生物医学进展》
CAS
2015年第14期2645-2648,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(31300706)
南京军区医学科技创新课题资助项目(11MA131)