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Capsule staining method for streptococcus pneumoniae

A Streptococcus pneumoniae and capsule staining technology, which is applied in the field of Streptococcus pneumoniae capsule staining, can solve the problems of high capsule shape requirements, high operation skills requirements, and high water content of bacterial capsules, and achieve high-efficiency strains The effect of low identification, skill and proficiency, and simple methods

Inactive Publication Date: 2015-06-17
BEIJING HUAAN INNOVATION BIOTECH
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  • Abstract
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AI Technical Summary

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

Method used

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  • Capsule staining method for streptococcus pneumoniae
  • Capsule staining method for streptococcus pneumoniae
  • Capsule staining method for streptococcus pneumoniae

Examples

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Effect test

Embodiment 1

[0051] Embodiment 1, the effect identification of Streptococcus pneumoniae capsule staining method

[0052] The present invention selects 7 serotype strains (table 1) in the 23-valent pneumococcal polysaccharide vaccine to carry out the effect display of the Streptococcus pneumoniae capsule staining method, and the steps are as follows:

[0053] 1. Preparation of bacterial specimens:

[0054] Take the lyophilized strain of Streptococcus pneumoniae, add 200 μl of sterile saline to dissolve, and repeatedly blow and beat to mix. Take 100 μl on 10% sheep blood MH medium, and spread evenly with a glass triangular applicator. placed in 6% CO 2 In the incubator, cultured upside down at 37°C for 16h-18h, washed with physiological saline, and smeared for later use.

[0055] 2. Smear: Take a small amount of prepared bacterial specimen and apply it evenly on a clean glass slide, scrape it into a thin layer, and let it dry naturally (because the water content of the capsule is above 90...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/14C12R1/46
Inventor 刘荷中赵泽坤翟振华
Owner BEIJING HUAAN INNOVATION BIOTECH
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