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通过噬菌体聚合酶链反应检测婴儿配方奶粉中存活的副结核分枝杆菌亚种,并通过培养进行确认。

Detection of viable Mycobacterium avium subspecies paratuberculosis in powdered infant formula by phage-PCR and confirmed by culture.

作者信息

Botsaris George, Swift Benjamin M C, Slana Iva, Liapi Maria, Christodoulou Maritsa, Hatzitofi Maria, Christodoulou Vasiliki, Rees Catherine E D

机构信息

Department of Agricultural Sciences, Biotechnology and Food Science, Cyprus University of Technology, Limassol, Cyprus.

Division of Food Sciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE12 5RD, United Kingdom.

出版信息

Int J Food Microbiol. 2016 Jan 4;216:91-4. doi: 10.1016/j.ijfoodmicro.2015.09.011. Epub 2015 Sep 21.

Abstract

Surveys from different parts of the world have reported that viable Mycobacterium avium subsp. paratuberculosis (MAP) can be cultured from approximately 2% of samples of retail pasteurised milk samples. Pasteurised milk is used for the production of powdered infant formula (PIF) and therefore there is a concern that MAP may also be present in these products. Several studies have previously reported the detection of MAP in PIF using PCR-based assays. However, culture-based surveys of PIF have not detected viable MAP. Here we describe a phage amplification assay coupled with PCR (page-PCR) that can rapidly detect viable MAP in PIF. The results of a small survey showed that the phage-PCR assay detected viable MAP in 13% (4/32) of PIF samples. Culture detected viable MAP in 9% (3/32) PIF samples, all of which were also phage-PCR positive. Direct IS900 PCR detected MAP DNA in 22% (7/32) of PIF samples. The presence of viable MAP in PIF indicates that MAP either survived PIF manufacturing or that post-production contamination occurred. Irrespective of the route of MAP contamination, the presence of viable MAP in PIF is a potential public health concern.

摘要

来自世界不同地区的调查显示,可从约2%的零售巴氏杀菌乳样本中培养出活的副结核分枝杆菌(MAP)。巴氏杀菌乳用于生产婴儿配方奶粉(PIF),因此人们担心这些产品中也可能存在MAP。此前有多项研究报告了使用基于PCR的检测方法在PIF中检测到MAP。然而,基于培养的PIF调查未检测到活的MAP。在此,我们描述了一种结合PCR的噬菌体扩增检测方法(噬菌体-PCR),该方法可快速检测PIF中的活MAP。一项小型调查结果显示,噬菌体-PCR检测方法在13%(4/32)的PIF样本中检测到活的MAP。培养法在9%(3/32)的PIF样本中检测到活的MAP,所有这些样本噬菌体-PCR检测也呈阳性。直接IS900 PCR在22%(7/32)的PIF样本中检测到MAP DNA。PIF中存在活的MAP表明MAP要么在PIF生产过程中存活下来,要么在生产后发生了污染。无论MAP污染的途径如何,PIF中存在活的MAP都是一个潜在的公共卫生问题。

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