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开发和优化一种环介导等温扩增(LAMP)检测方法,用于特异性检测扩展青霉。

Development and optimization of a loop-mediated isothermal amplification (LAMP) assay for the species-specific detection of Penicillium expansum.

机构信息

Technical University of Munich, TUM School of Life Sciences, Chair of Technical Microbiology, Gregor-Mendel-Str. 4, 85354, Freising, Germany.

Technical University of Munich, TUM School of Life Sciences, Chair of Technical Microbiology, Gregor-Mendel-Str. 4, 85354, Freising, Germany.

出版信息

Food Microbiol. 2021 May;95:103681. doi: 10.1016/j.fm.2020.103681. Epub 2020 Nov 16.

Abstract

Penicillium expansum is the main cause of Blue Mold Decay, which is the economically most significant postharvest disease on fruits. It occurs especially on pomaceous fruits such as apples and pears but also on a wide range of other fruits such as grapes or strawberries. Besides its negative economic effects on the industry, the fungus is also of health concern as it produces patulin, a mycotoxin known to provoke harmful effects in humans. A specific and rapid detection of this fungus therefore is required. In the current study, a loop-mediated isothermal amplification (LAMP) assay was developed and optimized for the species-specific detection of P. expansum. The assay showed high specificity during tests with genomic DNA of 187 fungal strains. The detection limit of the developed assay was 25 pg genomic DNA of P. expansum per reaction. The assay was successfully applied for the detection of the fungus on artificially contaminated apples, grapes, apple juice, apple puree, and grape juice. The developed assay is a promising tool for rapid, sensitive, specific, and cost-efficient detection of P. expansum in quality control applications in the food and beverage industry.

摘要

扩展青霉是青霉属的主要原因,它是果实采后最具经济意义的疾病。它尤其发生在苹果和梨等苹果属水果上,但也发生在葡萄或草莓等广泛的其他水果上。除了对该行业造成负面的经济影响外,该真菌还存在健康问题,因为它会产生展青霉素,一种已知会对人类产生有害影响的真菌毒素。因此,需要对这种真菌进行特异性和快速的检测。在当前的研究中,开发并优化了一种环介导等温扩增(LAMP)检测方法,用于青霉属物种的特异性检测。该方法在对 187 株真菌菌株的基因组 DNA 进行测试时表现出很高的特异性。该方法的检测限为每个反应 25pg 青霉属基因组 DNA。该方法成功地应用于人工污染的苹果、葡萄、苹果汁、苹果泥和葡萄汁中真菌的检测。该检测方法是一种很有前途的工具,可用于食品和饮料行业的质量控制应用中,快速、灵敏、特异性和经济高效地检测扩展青霉。

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