Wang Bei, Gan Qi, Tong Yue, Qiao Yanqi, Han Mengfan, Zhang Ruixiao, Han Quansheng, Li Chuanbing, Bai Su, Xu Long, Yin Yongmei, Zhang Chunze, Munkhtsetseg Baatar, Zhao Xiujie, Meng Meng, Xi Rimo
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and KLMDASR of Tianjin, Nankai University, Tongyan Road, Haihe Education Park, Tianjin 300350, China.
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and KLMDASR of Tianjin, Nankai University, Tongyan Road, Haihe Education Park, Tianjin 300350, China.
Talanta. 2023 May 1;256:124260. doi: 10.1016/j.talanta.2023.124260. Epub 2023 Jan 10.
Helicobacter pylori (H. pylori) infection has increasingly been a serious problem worldwide. The H. pylori infection can result in a series of stomach diseases including gastric carcinoma. There are two specific virulence genes (cagA and vacA) of H. pylori that are closely related to the occurrence of gastric cancer, and the common molecular detection methods (PCR, qPCR) are not suitable for high-screening test due to the requirement of expensive instruments and well-trained personals. Herein, we develop a rapid visual assay based on loop-mediated isothermal amplification (LAMP) for detecting H. pylori and its major virulence genes (cagA, vacAs1 and vacAm1) to guide clinical treatment for H. pylori infection. In this research, a fluorescent LAMP assay was established by optimizing the indicator of MnCl-Calcein, so that the resulted color and fluorescence changes could be utilized to perform the visual detection for H. pylori and its virulence genes with high sensitivity (10 ng/μL). The proposed LAMP assay is simple, fast (30 min) and capable in providing more sensitive results than traditional methods in the test of 46 clinical biopsy samples. By detecting the three virulence genes together, we can profile the infection risk of the patients, and discuss the correlation among the genes. Moreover, the method could be used to diagnose virulently infected individuals and benefit the eradication of H. pylori in early warning for gastric cancer.
幽门螺杆菌（H. pylori）感染在全球范围内日益成为一个严重问题。幽门螺杆菌感染可导致一系列胃部疾病，包括胃癌。幽门螺杆菌有两个与胃癌发生密切相关的特定毒力基因（cagA和vacA），而常用的分子检测方法（PCR、qPCR）由于需要昂贵的仪器和训练有素的人员，不适用于大规模筛查检测。在此，我们开发了一种基于环介导等温扩增（LAMP）的快速可视化检测方法，用于检测幽门螺杆菌及其主要毒力基因（cagA、vacAs1和vacAm1），以指导幽门螺杆菌感染的临床治疗。在本研究中，通过优化MnCl-钙黄绿素指示剂建立了一种荧光LAMP检测方法，从而利用产生的颜色和荧光变化对幽门螺杆菌及其毒力基因进行高灵敏度（10 ng/μL）的可视化检测。所提出的LAMP检测方法简单、快速（30分钟），并且在对46份临床活检样本的检测中能够提供比传统方法更灵敏的结果。通过同时检测这三个毒力基因，我们可以分析患者的感染风险，并探讨基因之间的相关性。此外，该方法可用于诊断毒力感染个体，有助于在胃癌早期预警中根除幽门螺杆菌。
