原料乳中沙门氏菌的快速过滤富集及PCR检测

Rapid collection of Salmonella in raw milk with filtration method and its PCR detection

尽管国内外以PCR技术为基础对食源性致病菌的检测手段已经基本成熟,但是由于大多数有8～12h的预增菌步骤,从而增加了检测时间和检测成本。建立了对原料乳中沙门氏菌的快速、简单、灵敏PCR检测技术。人工污染沙门氏菌的原乳,先离心脱脂,然后添加Na2EDTA获得澄清乳液,最后通过0.45μm微膜过滤收集菌体。整个PCR检测过程可在6.5h以内完成,且检测灵敏度可达到1～10mL-1。该方法值得推广,应用于原乳中该食源致病菌的日常检测。

Although many assays based on PCR for detecting foodborne pathogens have been established, most of those techniques require 8- 12 h pre-enrichment step and thus are time consuming. A rapid, simple and sensitive PCR assay was developed for the determination of Salmonella in raw milk here. The raw milk, artificially contaminated Salmonellae, was skimmed by centrifugation and disodium EDTA was added to get a clarified solution. The bacteria were collected with a 0.45 μm membrane filtration and then subjected to the PCR assay. The whole PCR detection can be finished in 6.5 h and reach 1-10 cuf/mL detection limit. The assay could deserve to be generalized and applied to routine detection for the foodbome pathogen in raw milk.