Clinical Laboratory Center, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Department of Gastroenterology, Emergency General Hospital, Beijing, China.
Diagn Microbiol Infect Dis. 2022 May;103(1):115661. doi: 10.1016/j.diagmicrobio.2022.115661. Epub 2022 Feb 23.
Infection with cytotoxin-associated gene A (cagA)-positive Helicobacter pylori (H. pylori) is associated with severe gastrointestinal disease. A rapid, simple, and convenient detection method for cagA-positive H. pylori was an urgent need. We have developed and evaluated a duplex recombinase aided amplification combined with lateral flow dipstick (Duplex RAA-LFD) assay for detection of cagA-positive H. pylori strains. The Duplex RAA-LFD successfully detected DNA extracts in 25 min at 39°C, with visual detection limits of 1.2 × 10 CFU/mL and 10 pg, respectively. No positive amplification was observed in 6 non-H. pylori strains, indicating higher specificity. When testing 56 clinical isolates, the sensitivity and specificity of the Duplex RAA-LFD assay were 96% and 100%, respectively, with Duplex PCR serving as the reference method. Therefore, the Duplex RAA-LFD assay is a potential rapid and effective alternative to detect cagA-positive H. pylori strains in fresh gastric mucosal tissue.
细胞毒素相关基因 A (cagA)-阳性幽门螺杆菌 (H. pylori) 的感染与严重的胃肠道疾病有关。因此，迫切需要一种快速、简单、方便的 cagA-阳性 H. pylori 检测方法。我们开发并评估了一种用于检测 cagA-阳性 H. pylori 菌株的双重组酶辅助扩增结合侧流层析法 (Duplex RAA-LFD) 检测方法。该 Duplex RAA-LFD 在 39°C 下 25 分钟内成功检测到 DNA 提取物，目视检测限分别为 1.2×10 CFU/mL 和 10 pg。在 6 株非 H. pylori 菌株中未观察到阳性扩增，表明具有更高的特异性。当测试 56 株临床分离株时，Duplex RAA-LFD 检测方法的灵敏度和特异性分别为 96%和 100%，Duplex PCR 作为参考方法。因此，Duplex RAA-LFD 检测方法是一种快速有效的替代方法，可用于检测新鲜胃黏膜组织中的 cagA-阳性 H. pylori 菌株。
