Nishikawa Y, Hase A, Ishii E, Kishi T
Department of Epidemiology, Osaka City Institute of Public Health and Environmental Sciences, Japan.
Appl Environ Microbiol. 1990 Jun;56(6):1547-50. doi: 10.1128/aem.56.6.1547-1550.1990.
A dot-blot, enzyme-linked immunosorbent method and a latex agglutination test were studied for their abilities to detect Vibrio cholerae serotype O1 in aquatic samples by testing artificially contaminated water as well as samples from natural potential sources. Water samples were preenriched with alkaline peptone and then enriched with Monsur peptone water. For the dot-blot test, enriched cultures of organisms in a small portion of the Monsur peptone water were transferred to a polyvinylidene difluoride membrane with a microfiltration apparatus. The enzyme-linked immunosorbent assay was performed by using biotin-labeled antibodies and avidin-biotin-peroxidase complex; brown dots developed in the wells that contained serotype O1 vibrios. Latex agglutination tests were performed by mixing 1 drop of the culture in Monsur with 1 drop of reagent coated with monoclonal antibody specific for antigen A. The sensitivities and specificities of the methods were compared with those of the colony-blot method, which identified individual colonies of V. cholerae O1 in mixed bacterial cultures on isolation media. Our results indicate that the dot-blot method is as sensitive as the colony-blot method and is useful for screening for V. cholerae serotype O1 even in specimens that are heavily contaminated with non-O1 vibrios.
通过检测人工污染水以及天然潜在来源的样本，研究了斑点印迹法、酶联免疫吸附法和乳胶凝集试验检测水生样本中霍乱弧菌O1血清型的能力。水样先用碱性蛋白胨预富集，然后用蒙苏尔蛋白胨水富集。对于斑点印迹试验，用微滤装置将一小部分蒙苏尔蛋白胨水中的富集培养物转移到聚偏二氟乙烯膜上。酶联免疫吸附测定通过使用生物素标记的抗体和抗生物素蛋白-生物素-过氧化物酶复合物进行；含有O1血清型弧菌的孔中出现棕色斑点。乳胶凝集试验通过将1滴蒙苏尔培养物与1滴包被有针对抗原A的单克隆抗体的试剂混合进行。将这些方法的敏感性和特异性与菌落印迹法进行比较，菌落印迹法可在分离培养基上鉴定混合细菌培养物中霍乱弧菌O1的单个菌落。我们的结果表明，斑点印迹法与菌落印迹法一样灵敏，即使在被非O1弧菌严重污染的标本中，也可用于筛选霍乱弧菌O1血清型。
