Gill Pooria, Alvandi Amir-Houshang, Abdul-Tehrani Hossein, Sadeghizadeh Majid
Department of Nanobiotechnology, Faculty of Basic Sciences, Tarbiat Modares University, Tehran 14115-175, Iran.
Diagn Microbiol Infect Dis. 2008 Oct;62(2):119-24. doi: 10.1016/j.diagmicrobio.2008.05.003. Epub 2008 Jul 3.
This study describes a nanodiagnostic method using thermophilic helicase-dependent isothermal amplification (tHDA) and gold nanoparticle probes for colorimetric detection of Helicobacter pylori DNA. The primers targeting ureC gene were used for the amplification of bacterial DNA by the isothermal tHDA reaction, resulting in the accumulation of DNA amplicons. The amplicons were hybridized with specific gold nanoparticle probes. The hybrids were colorimetrically detected by the assembly of gold nanoparticles. Using this method, we detected as little as 10 CFU mL(-1) of H. pylori within less than 1 h. Results obtained from the gastric biopsy samples showed 92.5% and 95.4% of sensitivity and specificity, respectively, in comparison with culture results, and 100% and 98.8% of sensitivity and specificity, respectively, in comparison with those of the histologic studies. Owing to its ease of operation, this assay significantly reduces the time and cost needed for the molecular diagnosis of H. pylori and has the potential to facilitate early detection of this pathogen.
本研究描述了一种纳米诊断方法，该方法利用嗜热解旋酶依赖性等温扩增（tHDA）和金纳米颗粒探针进行幽门螺杆菌DNA的比色检测。靶向ureC基因的引物用于通过等温tHDA反应扩增细菌DNA，从而导致DNA扩增子的积累。扩增子与特定的金纳米颗粒探针杂交。通过金纳米颗粒的组装对比色检测杂交体。使用该方法，我们在不到1小时内检测到低至10 CFU mL(-1)的幽门螺杆菌。与培养结果相比，胃活检样本获得的结果显示敏感性和特异性分别为92.5%和95.4%，与组织学研究结果相比，敏感性和特异性分别为100%和98.8%。由于其操作简便，该检测方法显著减少了幽门螺杆菌分子诊断所需的时间和成本，并有可能促进该病原体的早期检测。
