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首页> 外文期刊>Applied and Environmental Microbiology >rRNA Stability in Heat-Killed and UV-Irradiated EnterotoxigenicStaphylococcus aureus and Escherichia coliO157:H7
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rRNA Stability in Heat-Killed and UV-Irradiated EnterotoxigenicStaphylococcus aureus and Escherichia coliO157:H7

机译：rRNA在热杀死和紫外线辐射的肠毒素金黄色葡萄球菌和大肠杆菌O157：H7中的稳定性

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摘要

Differentiation of viable cells from nonviable cells is of considerable importance in the development of methods to detect foodborne pathogens. To study the suitability of 16S rRNA as an indicator of cell viability in nucleic acid-based detection assays, we examined rRNA stability in two representative foodborne pathogens,Escherichia coli O157:H7 and enterotoxigenicStaphylococcus aureus, which were inactivated by extreme heat, moderate heat, and UV irradiation. Cell death under all conditions was confirmed by a failure to grow in brain heart infusion broth after incubation for 48 h at 37°C. rRNA stability was monitored by a Northern blot analysis, and detection was evaluated by using reverse transcription (RT)-PCR performed with two primer sets (which produced 325- and 1,400-bp amplicons). rRNA of neither pathogen was detected by Northern blot analysis and RT-PCR after cells were killed by autoclaving at 121°C for 15 min. In contrast, intact rRNA of both pathogens were detected by Northern blotting and could be amplified by RT-PCR up to 48 h after cells were killed by heat treatment at 80°C and UV irradiation at 254 nm. rRNA was a suitable target molecule for monitoring bacterial viability under extreme heat conditions, but the presence of rRNA was not correlated with viability following moderate heat inactivation or UV irradiation of cells.
机译：在检测食源性病原体的方法的开发中，活细胞与非活细胞的分化非常重要。为了研究16S rRNA在基于核酸的检测方法中作为细胞生存能力指标的适用性，我们检查了两种代表性食源性病原体（大肠杆菌O157：H7和肠毒素金黄色葡萄球菌）的rRNA稳定性，这些病原体通过极热，中热，和紫外线照射。在所有条件下的细胞死亡均通过在37°C下孵育48小时后脑心输液肉汤无法生长来证实。通过RNA印迹分析监测rRNA的稳定性，并使用两个引物对（产生325和1,400 bp的扩增子）进行逆转录（RT）-PCR评估检测结果。通过在121°C高压灭菌15分钟杀死细胞后，通过Northern印迹分析和RT-PCR检测到两种病原体的rRNA。相比之下，两种病原体的完整rRNA可以通过Northern印迹法检测到，并且可以通过在80°C的热处理和254 nm的紫外线照射杀死细胞后48小时内通过RT-PCR扩增。 rRNA是监测极端高温条件下细菌生存力的合适靶分子，但在中等程度的热失活或细胞受到紫外线照射后，rRNA的存在与生存力无关。

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