Department of Health Inspection and Quarantine and Fujian Province Key Laboratory of Environment and Health, School of Public Health, Fujian Medical University, Fuzhou, Fujian, 350122, PR China.
Department of Epidemiology and Health Statistics and Key Laboratory of Ministry of Education for Gastrointestinal Cancer, School of Public Health, Fujian Medical University, Fuzhou, Fujian, 350122, PR China.
Talanta. 2022 Jun 1;243:123367. doi: 10.1016/j.talanta.2022.123367. Epub 2022 Mar 3.
Helicobacter pylori infections are threats to public health due to their high infection rate and drug resistance. Identification of single-nucleotide variants (SNVs) in H. pylori is crucial for both diagnosis and therapy. Yet the clinical testing of resistant H. pylori mutants is still facing some challenges, such as the selectivity is not good enough for SNVs in abundant wild-type DNA, the lack of clinical validation and the economical burden on patients. Herein, an X-shaped DNA probe with a toehold initiator was designed, which could specifically hybridize with certain genotype DNA due to the thermodynamically driven reaction. A competitive reaction was developed to amplify the thermodynamic difference between wild-type DNA and SNVs, diminishing the interference of wild-type DNA. By this means, multiple SNVs in H. pylori were successfully identified and two SNVs related to clarithromycin resistance are chosen as model targets. A paper strip was fabricated for visual, fast screening of SNVs. Furthermore, the approach was validated using clinical samples, and a point-of-care (POCT) testing diagnosis was executed on saliva samples, demonstrating its potential for the prevention and cure of H. pylori infections.
幽门螺杆菌感染因其高感染率和耐药性而成为公共卫生的威胁。鉴定幽门螺杆菌中的单核苷酸变异（SNVs）对于诊断和治疗都至关重要。然而，耐药幽门螺杆菌突变体的临床检测仍然面临一些挑战，例如对丰富的野生型 DNA 中的 SNVs 的选择性不够好，缺乏临床验证以及对患者的经济负担。本文设计了一种带有引发子的 X 形 DNA 探针，由于热力学驱动的反应，该探针可以特异性地与特定基因型的 DNA 杂交。开发了一种竞争反应来放大野生型 DNA 和 SNVs 之间的热力学差异，从而减少野生型 DNA 的干扰。通过这种方式，成功鉴定了幽门螺杆菌中的多个 SNVs，并选择了与克拉霉素耐药性相关的两个 SNVs 作为模型靶标。制作了一种纸条，用于进行 SNVs 的可视化、快速筛选。此外，该方法已通过临床样本进行了验证，并在唾液样本上进行了即时检测（POCT）测试诊断，表明其在预防和治疗幽门螺杆菌感染方面具有潜力。
