Test method for evaluating in-vitro antibacterial activity of antibacterial substance

A technology of antibacterial activity and antibacterial substances, applied in the field of biological control, can solve the problems of special properties of traditional Chinese medicine extracts, affecting the measurement of antibacterial zone, dark and turbid color, etc., and achieve the effect of intuitive and rapid antibacterial effect.

Active Publication Date: 2017-03-01
ANHUI UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the extract of traditional Chinese medicine has special properties, and its diffusion phenomenon in the agar plate is different from that of antibiotics. The in vitro antibacterial test of traditional Chinese medicine cannot completely copy the in vitro antibacterial test of antibiotics.
The main characteristics of Chinese medicine extracts are: (1) Dark and turbid color: After the drug diffuses, a darker and turbid drug diffusion circle appears in the agar. This circle coincides with the transparent circle where bacteria do not grow, which affects the measurement of the diameter of the inhibition zone. It is difficult to visually evaluate the antibacterial activity
like figure 2 Shown: In the two wells (01)(02) on the agar plate, the antibacterial phenomenon of the water extract of Scutellaria baicalensis with deep color and turbidity is diffused, wherein b1 is the dark brown traditional Chinese medicine diffusion circle (inner circle), and b2 is the light brown traditional Chinese medicine diffusion circle (outer circle). circle); (03) (04) two holes are colorless and clear antibiotic ceftriaxone sodium diffusion antibacterial phenomenon, wherein b3 is a transparent antibacterial zone, it can be seen that when the traditional plate agar diffusion method is used for antibacterial test, the antibiotic is colorless and clear , the drug diffusion zone does no

Method used

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  • Test method for evaluating in-vitro antibacterial activity of antibacterial substance
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  • Test method for evaluating in-vitro antibacterial activity of antibacterial substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Preparation of methylene blue agar plates

[0031] 1.1 Preparation of indicator bacteria suspension: Inoculate Salmonella on ordinary slant medium to activate and grow for 6-8 hours. Gently scrape the bacterial lawn on the slope with an inoculation loop and place it in sterile physiological saline, vortex shaker to fully shake and adjust the concentration of the bacterial suspension to 0.5 McFarland turbidity.

[0032] 1.2 Preparation of bacteria-containing methylene blue agar plate: Weigh methylene blue and add distilled water to prepare a 1% methylene blue aqueous solution, which is sterilized for use. After the M-H agar is sterilized in the Erlenmeyer flask, the water bath is kept at a constant temperature of 45°C. Add sterilized 1% methylene blue aqueous solution according to the ratio of adding 2ml methylene blue aqueous solution to every 100ml of bacteria-containing methylene blue agar. Add 0.5 McFarland turbidity bacterial suspension into 6ml of bacterial liq...

Embodiment 2

[0045] Application of methylene blue bacteria-containing agar plate diffusion method in external antibacterial test of traditional Chinese medicine extraction liquid

[0046] Test procedure: 1, the preparation method of the methylene blue bacteria-containing agar plate is the same as that of Example 1, and the strains are respectively 2 kinds of Salmonella enteritidis and Salmonella typhimurium;

[0047] 2. Use a puncher with a diameter of 6mm to punch holes on the methylene blue bacteria-containing agar plate, and add the Chinese medicine water extract into the holes for diffusion. The Chinese medicine water extract is respectively taken from the water extract of Schizandra chinensis and the extract of black plum, and the concentration is 1g. / mL.

[0048] In vitro antibacterial effect evaluation: After the plate was incubated at a constant temperature of 37°C for 18 hours, the diameter of the blue circle or the mixed color circle of the drug itself and the blue color on the ...

Embodiment 3

[0052] Antibacterial test of different concentrations of Scutellaria baicalensis water extract on methylene blue bacteria-containing agar plate diffusion method

[0053] Test procedure: 1, the methylene blue bacteria-containing agar plate preparation method is the same as in Example 1;

[0054] 2. Use a hole puncher with a diameter of 6mm to make holes on the methylene blue bacteria-containing agar plate, and add the water extract of Scutellaria baicalensis into the holes for diffusion. mL.

[0055] Evaluation of antibacterial effect in vitro: After the plate was incubated at a constant temperature of 37°C for 18 hours, the diameter of the blue circle or the mixed color circle of the drug itself and the blue color on the methylene blue agar plate was measured to determine the diameter of the inhibition zone and comprehensively evaluate the antibacterial activity.

[0056] Test results: if Figure 6 as shown, Figure 6 -a, 6-b, and 6-c are the results of the interaction betw...

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Abstract

The invention specifically discloses a test method for evaluating the in-vitro antibacterial activity of an antibacterial substance, belonging to the technical field of biological control. According to the method, a methylene blue agar plate containing bacteria is used for agar-plate-diffusion-process in-vitro antibacterial activity test. The method is visual and simple, realizes evaluation through observation of a colored inhibition zone and overcomes the disadvantage of difficulty in accurate measurement of the diameter of an inhibition zone due to deep and turbid colors and complex components during in-vitro antibacterial test of traditional Chinese medicines or other to-be-measured substances with similar properties by using traditional agar plate diffusion processes.

Description

technical field [0001] The invention relates to the technical field of biological control, in particular to a test method for evaluating antibacterial activity of antibacterial substances in vitro. Background technique [0002] In vitro antibacterial test is a test to determine the sensitivity of microorganisms to a certain substance in vitro. It is often used to evaluate the activity of inhibiting and killing pathogenic bacteria. It has been widely used in scientific research, production and clinical, such as screening of antibacterial drugs, tracking during extraction, Determination of antimicrobial spectrum, drug susceptibility test to guide clinical medication, etc. The agar plate diffusion method is one of the most commonly used methods at present. [0003] The traditional agar plate diffusion method is to spread the indicator bacteria on the surface of the agar or evenly mix it into the agar, and then use the paper method, Oxford cup method, punching method and other ...

Claims

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Application Information

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IPC IPC(8): C12Q1/18
CPCC12Q1/18C12Q2304/20
Inventor 田晔孔彦龙姚传福宋峰亮马慧慧许鹏程
Owner ANHUI UNIV OF SCI & TECH
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