Department of Clinical Laboratory, Hainan Provincial People's Hospital, Haikou 570311, Hainan, PR China.
Department of Clinical Research Center, Affiliated Hospital of Guizhou Medical University, Guiyang 550002, Guizhou, China.
J Nanosci Nanotechnol. 2020 Mar 1;20(3):1478-1485. doi: 10.1166/jnn.2020.17354.
The loop-mediated isothermal amplification (LAMP) assay as efficient and convenient detection method was applied to detect the vacuolating cytotoxin A () gene of Helicobacter pylori (). The extracted genomic DNA of , which was purified through magnetic nanoparticles (MNPs), was amplified through the LAMP reaction using designed primers. The effect of LAMP detected on gene was evaluated through agarose gel electrophoresis in a gel imaging system and fluorescence-intensity analysis after addition of fluorescent dye. 11 pathogenic bacterial strains of different species were found to be negative for , while only a single positive result was obtained for . The minimum detection limit of the gene was established as 100 fg. We used the primers with specificity and sensitivity, which were designed by the specificity analysis and sensitivity analysis system. Once developed, the LAMP assay was be used to the detection of the gene in the gastric juice of patients. In conclusion, the LAMP assay is an efficient and fast tool for detection of the gene, and also for direct detection of the gene in the gastric juice of patients, with high sensitivity and specificity. Most importantly, the proposed detection method shows promising potential for clinical application in the future, where it can greatly reduce the difficulty of detection and also shorten detection times.
环介导等温扩增(LAMP)检测法是一种高效、便捷的检测方法,被应用于检测空泡细胞毒素 A()基因的幽门螺杆菌()。提取的幽门螺杆菌基因组 DNA 通过磁纳米颗粒(MNPs)纯化,然后通过设计的引物进行 LAMP 反应扩增。通过琼脂糖凝胶电泳在凝胶成像系统中以及添加荧光染料后的荧光强度分析,评估 LAMP 对基因的检测效果。11 种不同种属的致病性细菌菌株均为基因阴性,而仅获得了单一的阳性结果。基因的最小检测限为 100 fg。我们使用了特异性和灵敏度分析系统设计的具有特异性和灵敏度的引物。一旦开发成功,LAMP 检测法将用于检测患者胃中的基因。总之,LAMP 检测法是一种高效、快速的基因检测工具,也可用于直接检测患者胃中的基因,具有高灵敏度和特异性。最重要的是,该检测方法具有广阔的临床应用前景,在未来有望极大地降低检测难度和缩短检测时间。