Saito N, Sato F, Kato M, Takeda H, Sugiyama T, Asaka M
The Third Department of Internal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
Helicobacter. 1998 Sep;3(3):170-3.
Helicobacter pylori (H. pylori) can be morphologically divided into spiral and coccoid forms. Although many different staining procedures for light microscopy have been developed to detect H. pylori, there are no reports that the coccoid forms can be specially identified by a staining method. The ability to identify the coccoid form of H. pylori by light microscopy would be important for studies evaluating its possible role in gastric disease. We examined whether both the coccoid and the spiral forms could be stained using anti-H. pylori antibody.
Specimens from the stomachs of dyspeptic patients with proven H. pylori infection and H. pylori from culture (1st and 4th day) were stained with the light microscopic immunogold silver enhancing stain using the rabbit IgG specific for H. pylori. Cultured H. pylori was also stained with electron microscopic immunogold staining method using the same antibody. The number of coccoid forms was counted by scanning electron microscopy.
Biopsies and H. pylori from 6 patients were studied. H. pylori from both biopsy material and culture appeared as black organisms by light microscopic immunogold silver enhancing stain. The coccoid forms constituted 0.4% and 98.3% on days 1 and 4 of culture, respectively (by scanning electron microscopy). The antigen recognized by the anti-H. pylori antibody was located on the surface of the flagella, the cell wall, or in the cytoplasm by immunoelectron microscopy.
This study shows that both spiral and coccoid forms of H. pylori can be detected by light microscopic immunogold silver enhancing stain using anti-H. pylori antibody. This simple stain is to be proven useful for studies aimed at understanding whether the coccoid form plays a role in disease.
幽门螺杆菌（H. pylori）在形态上可分为螺旋形和球形。尽管已开发出许多不同的光学显微镜染色方法来检测幽门螺杆菌，但尚无关于球形形态可通过染色方法进行特异性鉴定的报道。通过光学显微镜鉴定幽门螺杆菌的球形形态对于评估其在胃部疾病中可能作用的研究具有重要意义。我们研究了球形和螺旋形幽门螺杆菌是否都能用抗幽门螺杆菌抗体进行染色。
从确诊感染幽门螺杆菌的消化不良患者的胃中采集标本，以及培养（第1天和第4天）的幽门螺杆菌，使用对幽门螺杆菌特异的兔IgG，通过光学显微镜免疫金银增强染色法进行染色。培养的幽门螺杆菌还用相同抗体通过电子显微镜免疫金染色法进行染色。通过扫描电子显微镜计数球形形态的数量。
对6例患者的活检标本和幽门螺杆菌进行了研究。通过光学显微镜免疫金银增强染色法，活检材料和培养物中的幽门螺杆菌均呈现为黑色菌体。培养第1天和第4天，球形形态分别占0.4%和98.3%（通过扫描电子显微镜）。通过免疫电子显微镜观察，抗幽门螺杆菌抗体识别的抗原位于鞭毛表面、细胞壁或细胞质中。
本研究表明，使用抗幽门螺杆菌抗体通过光学显微镜免疫金银增强染色法可检测到幽门螺杆菌的螺旋形和球形形态。这种简单的染色方法有望对旨在了解球形形态是否在疾病中起作用的研究有用。
