基于链霉亲和素支架的 DNA 四联体的灵敏荧光 ELISA 法检测大肠杆菌 O157:H7。
Sensitive fluorescence ELISA with streptavidin scaffolded DNA tetrads for the detection of Escherichia coli O157:H7.
机构信息
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China.
College of Life Science, National R&D Center for Freshwater Fish Processing, Jiangxi Normal University, Nanchang 330022, China; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Province Center for Disease Control and Prevention, Nanchang 330029, China.
出版信息
J Dairy Sci. 2023 Sep;106(9):5930-5939. doi: 10.3168/jds.2022-23015. Epub 2023 Jul 18.
Escherichia coli O157:H7 poses a threat to humans. Traditional ELISA is not a sensitive method for the detection of E. coli O157:H7. Here, an efficient method was designed for improving the load capacity of alkaline phosphatase (ALP) with streptavidin scaffolded DNA tetrad (SS-DNAt). With more ALP, more ascorbic acid 2-phosphate was catalyzed to ascorbic acid that was used to synthesize fluorescence poly adenine-thymine-templated copper nanoclusters. Based on SS-DNAt, fluorescence ELISA was successfully proposed for improving the sensitivity for detection of E. coli O157:H7 in milk samples. The method showed a linear range of 10 to 10 cfu/mL. The limit of detection of fluorescence ELISA was 3.75 × 10 cfu/mL and 6.16-fold better than that of traditional ELISA. The recovery of the fluorescence ELISA was 86.7 to 93.6% with the coefficient of variation of 5.6 to 10.5% in milk. This method could be used to detect hazardous material in food.
大肠杆菌 O157:H7 对人类构成威胁。传统的 ELISA 不是检测大肠杆菌 O157:H7 的敏感方法。在这里，设计了一种有效的方法来提高链霉亲和素支架 DNA 四联体（SS-DNAt）的碱性磷酸酶（ALP）的负载能力。有了更多的 ALP，就会有更多的抗坏血酸 2-磷酸被催化成抗坏血酸，用于合成荧光多腺嘌呤-胸腺嘧啶模板铜纳米簇。基于 SS-DNAt，成功地提出了荧光 ELISA 方法，以提高牛奶样品中大肠杆菌 O157:H7 的检测灵敏度。该方法的线性范围为 10 至 10 cfu/mL。荧光 ELISA 的检测限为 3.75×10 cfu/mL，比传统 ELISA 提高了 6.16 倍。荧光 ELISA 在牛奶中的回收率为 86.7%至 93.6%，变异系数为 5.6%至 10.5%。该方法可用于检测食品中的有害物质。
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