基于荧光探针的重组酶辅助扩增快速检测大肠杆菌 O157:H7。
The fluorescent probe-based recombinase-aided amplification for rapid detection of Escherichia coli O157:H7.
机构信息
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China.
Ningbo International Travel Healthcare Center (Ningbo Customs Port Outpatient Department), Ningbo, 315010, PR China.
出版信息
Mol Cell Probes. 2021 Dec;60:101777. doi: 10.1016/j.mcp.2021.101777. Epub 2021 Nov 1.
Escherichia coli O157:H7 (E. coli O157:H7) is a common foodborne morbigenous microorganism, which can spread through fecal-oral transmission. Humans can be infected by ingesting foods and water contaminated with E. coli O157:H7, which can cause various symptoms. In present study, we have successfully developed a quick and hypersensitive fluorescent probe-based Recombinase-aided amplification (RAA) method and applied in E. coli O157:H7 detection at 39 °C in 20 min. The sensitivity of the assay in pure E. coli O157:H7 suspension was 5.6 × 10 CFU/mL. The fluorescent probe-based RAA assay was further applied in three samples, and the limit of detection (LOD) in skimmed milk, lettuces and lake water was 5.4 × 10 CFU/mL, 7.9 × 10 CFU/mL and 5.2 × 10 CFU/mL, separately. This method showed a high sensitivity and short detection time, which has the feasible application in on-site test in real samples.
大肠杆菌 O157:H7（E. coli O157:H7）是一种常见的食源性致病微生物，可通过粪-口途径传播。人类可以通过摄入被大肠杆菌 O157:H7 污染的食物和水而感染，从而导致各种症状。在本研究中，我们成功开发了一种快速、高灵敏度的荧光探针重组酶辅助扩增（RAA）方法，并应用于 39°C 下 20 分钟内检测大肠杆菌 O157:H7。在纯大肠杆菌 O157:H7 悬浮液中的检测灵敏度为 5.6×10 CFU/mL。该荧光探针 RAA 检测方法进一步应用于三种样品中，在脱脂乳、生菜和湖水的检测限（LOD）分别为 5.4×10 CFU/mL、7.9×10 CFU/mL 和 5.2×10 CFU/mL。该方法具有较高的灵敏度和较短的检测时间，可在现场实际样品中进行应用。
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