College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China.
Institute of Agro-Product Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
Biosensors (Basel). 2023 Apr 26;13(5):503. doi: 10.3390/bios13050503.
is one of four key global causes of diarrhea, and in humans, it is generally contracted through the consumption of contaminated food. It is necessary to develop an accurate, simple, and rapid method to monitor in the early phase. Herein, we developed a sequence-specific visualization method based on loop-mediated isothermal amplification (LAMP) for the detection of in milk. With restriction endonuclease and nicking endonuclease, amplicons were produced into single-stranded triggers, which further promoted the generation of a G-quadruplex by a DNA machine. The G-quadruplex DNAzyme possesses peroxidase-like activity and catalyzes the color development of 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) as the readouts. The feasibility for real samples analysis was also confirmed with spiked milk, and the sensitivity was 800 CFU/mL when observed with the naked eye. Using this method, the detection of in milk can be completed within 1.5 h. Without the involvement of any sophisticated instrument, this specific colorimetric method can be a useful tool in resource-limited areas.
志贺氏菌是全球四大腹泻病原菌之一，人类通常通过食用被污染的食物感染志贺氏菌。因此，有必要开发一种准确、简单、快速的方法来监测早期的志贺氏菌。本研究开发了一种基于环介导等温扩增（LAMP）的序列特异性可视化方法来检测牛奶中的志贺氏菌。通过限制内切酶和切口酶，扩增产物被切割成单链触发物，进一步通过 DNA 机器生成 G-四链体。G-四链体 DNA 酶具有过氧化物酶样活性，可催化 2,2'-联氮-双(3-乙基苯并噻唑啉-6-磺酸)（ABTS）显色作为读取信号。用添加志贺氏菌的牛奶验证了实际样品分析的可行性，肉眼观察的最低检测限为 800 CFU/mL。使用该方法，可在 1.5 小时内完成牛奶中志贺氏菌的检测。这种特异性比色法不需要任何复杂的仪器，可作为资源有限地区的有用工具。
