检出限
金黄色葡萄球菌
环介导等温扩增
荧光染料
化学
荧光
食品科学
细菌
微生物学
色谱法
实时聚合酶链反应
DNA
生物
生物化学
量子力学
基因
物理
遗传学
作者
Xingxing Chen,Xiaoli Wu,Min Gan,Feng Xu,Lihua He,Dong Yang,Hengyi Xu,Nagendra P. Shah,Hua Wei
标识
DOI:10.3168/jds.2014-8828
摘要
Staphylococcus aureus is one of the main pathogens in dairy and meat products; therefore, developing a highly sensitive and rapid method for its detection is necessary. In this study, a quantitative detection method for Staph. aureus was developed using silica-coated magnetic nanoparticles and thermophilic helicase-dependent isothermal amplification. First, genomic DNA was extracted from lysed bacteria using silica-coated magnetic nanoparticles and amplified using thermophilic helicase-dependent isothermal amplification. After adding the nucleic-acid dye SYBR Green I to the amplicons, the fluorescence intensity was observed using a UV lamp or recorded using a fluorescence spectrophotometer. This detection system had a detection limit of 5 × 100 cfu/mL in pure culture and milk-powder samples and 5 × 101 cfu/mL in pork samples using a UV light in less than 2 h. In addition, a good linear relationship was obtained between fluorescence intensity and bacterial concentrations ranging from 102 to 104 cfu/mL under optimal conditions. Furthermore, the results from contaminated milk powder and pork samples suggested that the detection system could be used for the quantitative analysis of Staph. aureus and applied potentially to the food industry for the detection of this pathogen.
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