Jiangsu Key Laboratory of Marine Bioresources and Environment, Huaihai Institute of Technology, Lianyungang, China.
Jiangsu Marine Resources Development Research Institute, Lianyungang, China.
Helicobacter. 2019 Jun;24(3):e12577. doi: 10.1111/hel.12577. Epub 2019 Apr 4.
Helicobacter pylori (H pylori) is a disease-causing pathogen capable of surviving under acidic conditions of the human stomach. Almost half of the world's population is infected with H pylori, with gastric cancer being the most unsatisfactory prognosis. Although H pylori has been discovered 30 years ago, the effective treatment and elimination of H pylori continue to be problematic.
In our study, we screened nucleic acid aptamers using H pylori surface recombinant antigens as targets. Trypsin was used for separating aptamers that were bound to proteins. Following nine rounds of screening, we performed sequence similarity analyses to assess whether the aptamers can recognize the target protein. Two sequences with desirable recognition ability were selected for affinity detection. Aptamer Hp4 with the strongest binding ability to the H pylori surface recombinant antigen was chosen. After optimization of the binding conditions, we conducted specificity tests for Hp4 using Escherichia coli, Staphylococcus aureus, Vibrioanguillarum, and H pylori.
The data indicated that the aptamer Hp4 had an equilibrium dissociation constant (K ) of 26.48 ± 5.72 nmol/L to the target protein. This aptamer was capable of exclusively detecting H pylori cells, without displaying any specificity for other bacteria.
We obtained a high-affinity aptamer for H pylori, which is expected to serve as a new molecular probe for detection of H pylori.
幽门螺杆菌（H.pylori）是一种能够在胃酸环境下生存的致病病原体。全球近一半的人口感染了 H.pylori，而胃癌是预后最不理想的疾病。尽管 H.pylori 早在 30 年前就已被发现，但有效治疗和根除 H.pylori 仍存在问题。
在本研究中，我们以 H.pylori 表面重组抗原为靶标，筛选核酸适配体。采用胰蛋白酶分离与蛋白结合的适配体。经过九轮筛选，我们进行序列相似性分析，以评估适配体是否能识别目标蛋白。选择具有良好识别能力的两条序列进行亲和力检测。选择与 H.pylori 表面重组抗原结合能力最强的适配体 Hp4 进行优化。优化结合条件后，我们使用大肠杆菌、金黄色葡萄球菌、鳗弧菌和 H.pylori 对 Hp4 进行特异性测试。
数据表明，适配体 Hp4 与靶蛋白的平衡解离常数（K ）为 26.48±5.72nmol/L。该适配体能够特异性地检测 H.pylori 细胞，而对其他细菌没有特异性。
我们获得了一种对 H.pylori 具有高亲和力的适配体，有望成为检测 H.pylori 的新型分子探针。
