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Rapid Methods for Quality Assurance of Foods: the Next Decade with Polymerase Chain Reaction (PCR)-Based Food Monitoring

机译:食品质量保证的快速方法:基于聚合酶链反应(PCR)的食品监测的下一个十年

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摘要

Microbiological analysis is an integral part of food quality control, as well as of the management of food chain safety. Microbiological testing of foodstuffs complements the preventive approach to food safety activities based mainly on implementation and application of the concept of Hazard Analysis and Critical Control Points (HACCP). Traditional microbiological methods are powerful but lengthy and cumbersome and therefore not fully compatible with current requirements. Even more, pathogens exist that are fastidious to cultivate or uncultivable at all. Besides immunological tests, molecular methods, specifically those based on polymerase chain reaction (PCR), are available options to meet industry and enforcement needs. The clear advantage of PCR over all other rapid methods is the striking analytical principle that is based on amplification of DNA, a molecule being present in every cell prone to multiply. Just by changing primers and probes, different genomes such as bacteria, viruses or parasites can be detected. A second advantage is the ability to both detect and quantify a biotic contaminant. Some previously identified obstacles of implementation of molecular methods have already been overcome. Technical measures became available that improved robustness of molecular methods, and equipment and biochemicals became much more affordable. Unfortunately, molecular methods suffer from certain drawbacks that hamper their full integration to food safety control. Those encompass a suitable sample pre-treatment especially for a quantitative extraction of bacteria and viruses from solid foods, limited availability of appropriate controls to evaluate the effectiveness of the analytical procedure, the current inability of molecular methods to distinguish DNA from viable cells and DNA from dead or non-cultivable cells, and the slow progress of international harmonisation and standardisation, which limit full acceptance of PCR-based methods in food control. The aim of this review is to describe the context and the prospects of PCR-based methods, as well as trends in research and development aimed at solving the next decade challenges in order to achieve full integration of molecular methods in food safety control.
机译:微生物分析是食品质量控制以及食品链安全管理不可或缺的一部分。食品的微生物检测主要是基于危害分析和关键控制点(HACCP)概念的实施和应用,补充了食品安全活动的预防方法。传统的微生物学方法功能强大,但又冗长又麻烦,因此与当前要求不完全兼容。更甚者,存在着难以培养或根本无法耕种的病原体。除免疫学测试外,分子方法,尤其是基于聚合酶链反应(PCR)的分子方法,可满足工业和执法需求。与其他所有快速方法相比,PCR的明显优势是基于DNA扩增的惊人分析原理,该分子存在于每个易于繁殖的细胞中。仅通过改变引物和探针,就可以检测出不同的基因组,例如细菌,病毒或寄生虫。第二个优点是能够检测和定量生物污染物。已经克服了一些先前确定的实施分子方法的障碍。可以采用技术措施来提高分子方法的鲁棒性,并且设备和生化试剂的价格也变得更加便宜。不幸的是,分子方法具有某些缺点,妨碍了它们完全整合到食品安全控制中。这些措施包括适当的样品预处理,特别是用于从固体食品中定量提取细菌和病毒,有限的适当对照来评估分析程序的有效性,目前分子方法无法区分活细胞和活细胞DNA。死细胞或不可培养细胞,以及国际协调和标准化进展缓慢,这限制了在食品控制中完全接受基于PCR的方法。这篇综述的目的是描述基于PCR的方法的背景和前景,以及旨在解决下一个十年挑战的研究和发展趋势,以便将分子方法完全整合到食品安全控制中。

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