Rapid Detection of Helicobacter pylori by the Naked Eye Using DNA Aptamers,ACS Omega

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Rapid Detection of Helicobacter pylori by the Naked Eye Using DNA Aptamers
ACS Omega ( IF 3.7 ) Pub Date : 2021-01-21 , DOI: 10.1021/acsomega.0c05374
Hangjie Wu _{1,

2} , Lide Gu _{1,

2} , Xiaoyi Ma _{1,

2} , Xueqing Tian _{1,

2} , Shihui Fan _{1,

2} , Mingcan Qin _{1,

2} , Jing Lu _{1,

2} , Mingsheng Lyu _{1,

2} , Shujun Wang _{1,

2}

Affiliation

Helicobacter pylori was first isolated from gastritis patients by Barry J. Marshall and J. Robin Warren in 1982, and more than 90% of duodenal ulcers and about 80% of gastric ulcers are caused by H. pylori infection. Most detection methods require sophisticated instruments and professional operators, making detection slow and expensive. Therefore, it is critical to develop a simple, fast, highly specific, and practical strategy for the detection of H. pylori. In this study, we used H. pylori as a target to select unique aptamers that can be used for the detection of H. pylori. In our study, we used random ssDNA as an initial library to screen nucleic acid aptamers for H. pylori. We used binding rate and the fluorescence intensity to identify candidate aptamers. One DNA aptamer, named HPA-2, was discovered through six rounds of positive selection and three rounds of negative selection, and it had the highest affinity constant of all aptamers tested (K_d = 19.3 ± 3.2 nM). This aptamer could be used to detect H. pylori and showed no specificity for other bacteria. Moreover, we developed a new sensor to detect H. pylori with the naked eye for 5 min using illumination from a hand-held flashlight. Our study provides a framework for the development of other aptamer-based methods for the rapid detection of pathogenic bacteria.

中文翻译：

使用DNA适配子通过裸眼快速检测幽门螺杆菌

幽门螺杆菌最早是由Barry J. Marshall和J. Robin Warren于1982年从胃炎患者中分离出来的，超过90％的十二指肠溃疡和约80％的胃溃疡是由幽门螺杆菌感染引起的。大多数检测方法需要复杂的仪器和专业的操作人员，这使得检测速度缓慢且昂贵。因此，开发简单，快速，高度特异性和实用的幽门螺杆菌检测策略至关重要。在这项研究中，我们使用幽门螺杆菌作为靶标，以选择可用于检测幽门螺杆菌的独特适体。在我们的研究中，我们使用随机ssDNA作为初始文库来筛选幽门螺杆菌的核酸适体。我们使用结合率和荧光强度来鉴定候选适体。通过六轮正选择和三轮负选择发现了一种名为HPA-2的DNA适体，在所有测试的适体中，它具有最高的亲和常数（K _d = 19.3±3.2 nM）。该适体可用于检测幽门螺杆菌，对其他细菌没有特异性。此外，我们开发了一种新型传感器，可使用手持手电筒的照明用肉眼检测幽门螺杆菌5分钟。我们的研究为快速检测病原细菌的其他基于适体的方法提供了框架。

更新日期：2021-02-09

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