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qPCR for quantitative detection of tyramine-producing bacteria in dairy products

机译:qPCR定量检测乳制品中产生酪胺的细菌

摘要

Biogenic amines (BAs) are organic nitrogenous compounds that occur naturally in small concentrations in most living organisms, in which they have different biological functions. However, they can accumulate in foods due to the decarboxylating activity of certain bacteria. The consumption of food containing large amounts of some BAs can have toxicological consequences. Tyramine is one of the most active and common BAs found in cheeses. This article reports the design of an real-time quantitative PCR (qPCR) qPCR method for the detection and quantification of the microorganisms responsible for tyramine production in cheeses. A good linear relationship was obtained between the quantity of tyramine-producer DNA used in PCR reactions and the Ct (threshold cycle) value. The method was validated using 57 cheese samples in which the tyramine concentration was analysed by HPLC. Tyramine-producing bacteria were detected in different numbers in all samples, although tyramine itself was only detected in 56.1% of the samples. The results suggest that, independent of technological conditions such as pasteurisation or the ripening period, the presence of tyramine-producing bacteria over a threshold limit of 104 cfu g-1 - easily detected by the proposed qPCR method - leads to the accumulation of tyramine beyond recommendable limits. © 2009 Elsevier Ltd. All rights reserved.
机译:生物胺(BAs)是一种有机含氮化合物,在大多数活生物体中以低浓度天然存在,它们具有不同的生物学功能。但是,由于某些细菌的脱羧活性,它们会在食物中积累。食用含有大量某些BA的​​食物会产生毒理学后果。酪胺是奶酪中发现的最活跃,最常见的BA。本文报告了一种实时定量PCR(qPCR)qPCR方法的设计,该方法用于检测和定量负责奶酪中酪胺产生的微生物。在PCR反应中使用的酪胺生成DNA的数量与Ct(阈值循环)值之间获得了良好的线性关系。使用57个干酪样品对方法进行了验证,其中通过HPLC分析了酪胺浓度。尽管仅在56.1%的样品中检测到了酪胺本身,但在所有样品中均检测到了不同数量的产生酪胺的细菌。结果表明,与巴氏灭菌或成熟期等技术条件无关,酪氨酸生产细菌的存在超过104 cfu g-1的阈值极限-通过建议的qPCR方法很容易检测到-导致酪胺的积累超过推荐的限制。 ©2009 ElsevierLtd。保留所有权利。

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