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一种基于新型智能手机的比色适体传感器,用于现场检测牛奶中的大肠杆菌 O157:H7。

A novel smartphone-based colorimetric aptasensor for on-site detection of Escherichia coli O157:H7 in milk.

机构信息

Key Laboratory of Dairy Science, Ministry of Education, College of Food Science, Northeast Agricultural University, Harbin, China, 150030.

Key Laboratory of Dairy Science, Ministry of Education, College of Food Science, Northeast Agricultural University, Harbin, China, 150030.

出版信息

J Dairy Sci. 2021 Aug;104(8):8506-8516. doi: 10.3168/jds.2020-19905. Epub 2021 May 28.

Abstract

Effective testing tools for Escherichia coli O157:H7 can prevent outbreaks of foodborne illness. In this paper, a smartphone-based colorimetric aptasensor was developed using functionalized gold nanoparticles (GNP) and multi-walled carbon nanotubes (MWCNT) for monitoring E. coli O157:H7 in milk. The maximum absorption peak of GNP bonded with aptamer (Apt) generated evident transformation from 518 to 524 nm. The excess GNP-Apt was removed by functionalized MWCNT magnetized with carbonyl iron powder (CIP) and hybridized with a DNA probe, whereas the GNP-Apt immobilized on E. coli O157:H7 remained in the system. In the presence of a high-salt solution, the GNP-Apt that captured E. coli O157:H7 remained red, but the free GNP-Apt aggregated and appeared blue. The chromogenic results were analyzed by a smartphone-based colorimetric device that was fabricated using acrylic plates, a light-emitting diode, and a mobile power pack. To our knowledge, this was the first attempt to use a smartphone-based colorimetric aptasensor employing the capture of GNP-Apt coupled with separation of MWCNT@CIP probe to detect E. coli O157:H7. The aptasensor exhibited good reproducibility and no cross-reaction for other bacteria. A concentration of 8.43 × 10 cfu/mL of E. coli O157:H7 could be tested in pure culture, and 5.24 × 10 cfu/mL of E. coli O157:H7 could be detected in artificially contaminated milk after 1 h of incubation. Therefore, the smartphone-based colorimetric aptasensor was an efficient tool for the detection of E. coli O157:H7 in milk.

摘要

用于检测大肠杆菌 O157:H7 的有效测试工具可以预防食源性疾病的爆发。本文开发了一种基于智能手机的比色适体传感器,该传感器使用功能化金纳米粒子(GNP)和多壁碳纳米管(MWCNT)监测牛奶中的大肠杆菌 O157:H7。与适配体(Apt)结合的 GNP 的最大吸收峰从 518nm 显著转变为 524nm。多余的 GNP-Apt 通过被羰基铁粉(CIP)磁化的功能化 MWCNT 去除,并与 DNA 探针杂交,而与大肠杆菌 O157:H7 结合的 GNP-Apt 则留在系统中。在高盐溶液存在的情况下,捕获了大肠杆菌 O157:H7 的 GNP-Apt 保持红色,但游离的 GNP-Apt 聚集并呈现蓝色。比色结果通过使用亚克力板、发光二极管和移动电源包制造的基于智能手机的比色设备进行分析。据我们所知,这是首次尝试使用基于智能手机的比色适体传感器,该传感器采用捕获 GNP-Apt 并结合分离 MWCNT@CIP 探针来检测大肠杆菌 O157:H7。该适体传感器表现出良好的重现性,对其他细菌没有交叉反应。在纯培养物中,可以检测到浓度为 8.43×10 cfu/mL 的大肠杆菌 O157:H7,而在孵育 1 小时后,在人工污染的牛奶中可以检测到 5.24×10 cfu/mL 的大肠杆菌 O157:H7。因此,基于智能手机的比色适体传感器是一种检测牛奶中大肠杆菌 O157:H7 的有效工具。

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