D'iakov S I, Lebedeva I K, Lisin V V, Grishin G I
Antibiotiki. 1982 Oct;27(10):761-6.
A new procedure for rapid determination of the levels of antibiotic sensitivity in pathogenic microorganisms with the use of fluorescent antibodies is described. The procedure was developed with the use of a model of the vaccinal strains of Bacillus anthracis. It is based on determination of the microbial antibiotic resistance with the method of serial dilutions on solid media. Still, the medium with an antibiotic is inoculated instead of the pathogen with the native material subject to the analysis. The antibiotic effect on the microorganism is estimated with the method of fluorescent antibodies. The replica preparations obtained as a result of the pathogen growth in a mixed culture on nutrient media containing definite concentrations of the antibiotic are examined with the method of luminescence microscopy. The modification of the immunofluorescent procedure for rapid determination of the microbial sensitivity to antibiotics does not require obligatory isolation of the pathogen as a pure culture. This makes the procedure more economic with respect to the time necessary for the analysis. The following conditions for performing rapid analysis with respect to Bacillus anthracis are required: the minimal concentration of the pathogen in the specimen (2.10(5) spores/ml), preliminary thermal treatment of the specimen for destroying the spore microflora, additional cultivation for 6-8 hours at 37 degrees C. The presence of the accompanying sporulating microflora, i.e. common microorganisms present in the atmosphere, soil and open water bodies does not prevent the performance of the analysis.
描述了一种使用荧光抗体快速测定病原微生物抗生素敏感性水平的新方法。该方法是利用炭疽芽孢杆菌疫苗株模型开发的。它基于在固体培养基上通过连续稀释法测定微生物的抗生素抗性。然而,接种的是含有抗生素的培养基,而不是用待分析的天然材料接种病原体。用荧光抗体法评估抗生素对微生物的作用。对在含有一定浓度抗生素的营养培养基上混合培养中病原体生长所得到的复制制剂,用发光显微镜法进行检查。用于快速测定微生物对抗生素敏感性的免疫荧光方法的改进,不要求必须将病原体分离为纯培养物。这使得该方法在分析所需时间方面更经济。对炭疽芽孢杆菌进行快速分析需要以下条件:标本中病原体的最低浓度(2×10⁵ 个孢子/毫升),对标本进行初步热处理以破坏孢子微生物群落,在37℃下额外培养6 - 8小时。伴随的产孢微生物群落的存在,即大气、土壤和开放水体中存在的常见微生物,并不妨碍分析的进行。