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用于检测食品样本中单核细胞增生李斯特菌的活性PCR方法的优化

Optimization of a Viability PCR Method for the Detection of Listeria monocytogenes in Food Samples.

作者信息

Agustí Gemma, Fittipaldi Mariana, Codony Francesc

机构信息

GenIUL, Carrer de la Ciutat d'Assunción 4, 08030, Barcelona, Spain.

Laboratori de Microbiologia Sanitària i Mediambiental (MSM-Lab), Universitat Politècnica de Catalunya, Edifici GAIA, Rambla Sant Nebridi, 22, 08222, Terrassa, Barcelona, Spain.

出版信息

Curr Microbiol. 2018 Jun;75(6):779-785. doi: 10.1007/s00284-018-1448-6. Epub 2018 Feb 12.

DOI:10.1007/s00284-018-1448-6
PMID:29435612
Abstract

Rapid detection of Listeria and other microbial pathogens in food is an essential part of quality control and it is critical for ensuring the safety of consumers. Culture-based methods for detecting foodborne pathogens are time-consuming, laborious and cannot detect viable but non-culturable microorganism, whereas viability PCR methodology provides quick results; it is able to detect viable but non-culturable cells, and allows for easier handling of large amount of samples. Although the most critical point to use viability PCR technique is achieving the complete exclusion of dead cell amplification signals, many improvements are being introduced to overcome this. In the present work, the yield of dead cell DNA neutralization was enhanced by incorporating two new sample treatment strategies: tube change combined with a double light treatment. This procedure was successfully tested using artificially contaminated food samples, showing improved neutralization of dead cell DNA.

摘要

快速检测食品中的李斯特菌和其他微生物病原体是质量控制的重要组成部分,对于确保消费者安全至关重要。基于培养的食源性病原体检测方法耗时、费力,且无法检测活的但不可培养的微生物,而活力PCR方法能快速得出结果;它能够检测活的但不可培养的细胞,并且便于处理大量样本。尽管使用活力PCR技术的最关键之处在于完全排除死细胞扩增信号,但目前正在引入许多改进措施来克服这一问题。在本研究中,通过采用两种新的样品处理策略:换管结合双重光照处理,提高了死细胞DNA的中和效率。该方法在人工污染的食品样本中成功进行了测试,显示出对死细胞DNA的中和效果有所改善。

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