Capsule staining method for streptococcus pneumoniae

Abstract

The invention discloses a capsule staining method for streptococcus pneumonia, and relates to the field of microbiological detection. The capsule staining method comprises the following steps: (1) preparing a bacterium specimen; (2) performing smearing, namely uniformly smearing a clean slide with a small amount of bacterium specimen, scraping the bacterium specimen into a thin layer, and naturally air-drying the slide; (3) performing staining: namely adding dropwise 100 to 150 microliters of carbolic acid fuchsin staining liquor onto the slide to uniformly cover the slide, and naturally air-drying the slide; (4) performing mordanting, namely washing the slide, naturally air-drying the slide, adding dropwise a mordant to uniformly cover the slide for 3 to 5min, rinsing the slide with water, and naturally air-drying the slide; (5) performing microscopic examination. The capsule staining method for streptococcus pneumonia is simple, easy, low in time consumption and high in success rate and repeatability, and has excellent application prospect in the clinical diagnosis of pneumonia, and strains of streptococcus pneumonia can be rapidly and efficiently identified.

Description

technical field [0001] The invention relates to the field of microorganism detection, in particular to a capsule staining method of Streptococcus pneumoniae. Background technique [0002] Capsule is a mucus or colloidal substance secreted by bacteria under certain conditions. It is usually composed of polysaccharides, polypeptides or polysaccharide-protein complexes. Different bacteria have different capsule components. The capsule of Streptococcus pneumoniae is mainly composed of polysaccharides, which can be divided into more than 90 serotypes according to the different components of the capsule (Ding Shaoqing, Ye Renbang, Yuan Zenglin. Research on the serotyping of Streptococcus pneumoniae. Journal of Biological Products. 1988, 1(1):18-22), and different types of serotypes have different capsule thickness. Capsule is a special structure of bacteria. The low refractivity of capsule and poor ability of affinity dyes make it difficult to color. Making a good capsule stained...

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Problems solved by technology

[0018] The above methods and some other methods have certain disadvantages: firstly, the operation process is relatively complicated, and the operator’s operating skills are required to be high, which may lead to experiment failure and poor repeatability; secondly, these methods can be seen clearly after dyeing. There are very few bacteria in the capsule, which needs to be searched in a large area under the microscope, which is inconvenient for actual operation; thirdly, due to the high water content of the bacterial capsule, heating will cause the bacteria to lose water and shrink, and they will separate from the dye around the cells to produce a transparent bright color. area, causing false positives to appear

In addition, some improved capsule staining methods have higher requirements on the morphology of the capsule. In order to enhance the staining effect of the capsule in the process of preparing bacterial slices, it is necessary to add capsule protection solution, which is cumbersome and time-consuming, and the success rate is low. low, poor repeatability

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