Method for testing bacterial drug sensitivity by TTC (2,3,5-triphenyte-trazoliumchloride) reduction reaction

Abstract

The invention provides a method for testing bacterial drug sensitivity by TTC (2,3,5-triphenyte-trazoliumchloride) reduction reaction. The method comprises the steps as follows: preparation of an antibiotic solution; preparation of a 96-well plate with gradient antibiotic concentration; preparation of a bacterial suspension; constant temperature culture; result interpretation: if a medium is red after color reaction, the tested bacteria can tolerate antibiotics at the concentration, drug sensitivity test results are judged sensitively and accurately, the minimum inhibitory concentration is obtained, and thus, errors caused by visual judgment of operators and errors produced by turbidity of the medium when an instrument reads optical density value of a culture in plate wells are avoided. Then, in combination with CLSI (the Clinical Laboratory Standard Institute) identification standards, the sensitivity results of the tested bacteria to various antibiotics are interpreted to provide a guidance basis for selection of reasonable antibiotic treatment, and thus, accurate evaluation results are provided for antibacterial spectra and antibacterial activity of new drugs, bacterial drug resistance is better monitored, the epidemic trend of drug-resistant bacteria is grasped, and food and public health safety are ensured.

Description

technical field [0001] The invention relates to a method for detecting bacterial drug sensitivity, in particular to a method for detecting bacterial drug sensitivity by utilizing TTC reduction reaction, and belongs to the field of microorganisms. Background technique [0002] The widespread use of antibiotics in animal husbandry, veterinary medicine and clinical treatment has led to the emergence of a large number of drug-resistant bacteria, which have become one of the major threats to global food safety and public health security. The detection and determination of drug-resistant bacteria is mainly achieved through drug sensitivity testing. The test results are also an important reference for rational selection of antibiotics, improvement of efficacy and avoidance of abuse of antibiotics in the clinical treatment of bacterial infectious diseases. It is also an important reference for the development of drug resistance mechanisms. basis. [0003] At present, the commonly u...

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Problems solved by technology

Agar dilution method: It can detect the drug susceptibility of a large number of strains at the same time, with good repeatability, but the workload is large, the amount of antibiotics and culture medium is used, and the drug sensitivity detection of a small number of strains is time-consuming and labor-intensive; the disc diffusion method: The operation is simple and widely used, but the test results will be affected by many factors such as the quality of the drug-sensitive paper and the type of culture medium, and the minimum inhibitory concentration of antibiotics on the tested bacteria cannot be detected, and the drug-sensitive paper cannot be prepared by itself; Micro-broth dilution method: it saves reagents and materials, but its biggest limitation is that it is difficult to accurately judge whether the bacteria in the wells of the plate are growing and the results of the minimum inhibitory concentration of antibiotics with the naked eye, and the results of drug susceptibility testing may vary depending on the test. Human judgment errors are caused by human visual errors, such as reading the optical density value of the culture in the plate well with a microplate reader or a spectrophotometer, which causes a large error due to the influence of the turbidity of the medium itself

It can be seen that several methods commonly used to detect bacterial drug susceptibility have certain limitations.

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