Skindersoe Mette E, Rasmussen Lone, Andersen Leif P, Krogfelt Karen A
Microbiology and Infection Control, Statens Serum Institute, Artillerivej 5, DK-2300, Copenhagen S, Denmark.
Department of Clinical Microbiology 9321, Copenhagen University Hospital, Rigshospitalet, Blegdamsvej 9, DK-2100, Copenhagen, Denmark.
Helicobacter. 2015 Jun;20(3):199-205. doi: 10.1111/hel.12191. Epub 2015 Jan 21.
Reducing adhesion of Helicobacter pylori to gastric epithelial cells could be a new way to counteract infections with this organism. We here present a novel method for quantification of Helicobacter pylori adhesion to cells.
Helicobacter pylori is allowed to adhere to AGS or MKN45g cells in a 96-well microtiter plate. Then wells are added saponin, which lyses the cells without affecting the bacteria. After addition of alamarBlue(®) (resazurin) and 1- to 2-hour incubation, fluorescence measurements can be used to quantify the number of adherent bacteria.
By use of the method, we demonstrate that adhesion of both a sabA and babA deletion mutant of H. pylori is significantly reduced compared to the wild type.
The method offers a number of applications and may be used to compare the adherence potential of different strains of H. pylori to either cells or different materials or to screen for potential anti-adhesive compounds. The results presented here suggest that this easy and reproducible assay is well suited for quantitative investigation of H. pylori adhesion.
降低幽门螺杆菌对胃上皮细胞的黏附可能是对抗该菌感染的一种新方法。我们在此介绍一种定量检测幽门螺杆菌对细胞黏附的新方法。
将幽门螺杆菌接种于96孔微量滴定板中的AGS或MKN45g细胞上。然后向孔中加入皂角苷，其可裂解细胞而不影响细菌。加入alamarBlue（刃天青）并孵育1至2小时后，可通过荧光测量来定量黏附细菌的数量。
通过使用该方法，我们证明幽门螺杆菌sabA和babA缺失突变体的黏附与野生型相比显著降低。
该方法有多种应用，可用于比较不同幽门螺杆菌菌株对细胞或不同材料的黏附潜力，或筛选潜在的抗黏附化合物。此处给出的结果表明，这种简便且可重复的检测方法非常适合用于幽门螺杆菌黏附的定量研究。
