Antibacterial drug sensitivity test detection method based on fluorescent D-type amino acid metabolism marker

Abstract

The invention relates to an antibacterial drug sensitivity test detection method based on a fluorescent D-type amino acid metabolism marker. According to the method, the fluorescent D-type amino acidmetabolism marker is utilized to mark a bacterial solution obtained by mixing and incubating with an antibiotic solution, so that the minimum inhibitory concentration of antibiotics can be obtained according to the fluorescence signal intensity of the bacterial solution.

Description

technical field [0001] The present invention relates to the evaluation method of bacterial metabolic activity and the detection method of antimicrobial susceptibility test, especially to the detection method of antimicrobial susceptibility test based on fluorescent D-type amino acid metabolism label, for rapid evaluation or evaluation of antibacterial agent or bacterial metabolic activity, for example Antibiotics, antibacterial effect on clinically isolated bacterial strains, to guide clinical medication. Background technique [0002] With the widespread use of antibiotics in medical and farming industries, especially the extensive use of broad-spectrum antibiotics, the problem of bacterial resistance has become increasingly prominent, and even super bacteria that can resist almost all antibiotics have emerged one after another. The emergence and spread of bacterial resistance has attracted worldwide attention, and is considered by the World Health Organization to be one of ...

AI Technical Summary

Problems solved by technology

For example, most microfluidic-based bacterial microscopic imaging technologies can directly observe the growth, proliferation or morphological changes of bacteria under the action of antibiotics. The amount of samples and reagents is greatly reduced, but microfluidic technology has not yet been popularized. The technical threshold for judging by microscopic imaging is high, the promotion is difficult, and the equipment cost is high. Moreover, based on the detection mechanism of bacterial growth, proliferation or morphological changes, it is impossible to identify the Bacteria in non-growing but metabolically active (NGMA) state, prone to misinterpretation [Refs 3, 4, 5]

The RNA detection method judges the sensitivity of bacteria to drugs by detecting the changes in the transcripts of bacteria under the action of antibiotics. Can determine whether the bacteria are sensitive to antibiotics, but cannot quantify the MIC [References 6, 7]

Although the atomic force microscope cantilever method judges the metabolic activity of bacteria by microscopic observation and monitoring the movement of bacteria under the action of antibiotics (detection of bacterial metabolic activity can effectively identify bacteria in a state of non-growth but metabolic activity (NGMA), which is more accurate , but this method is greatly affected by various factors (such as flowing liquid, bacterial quantity, etc.) during detection and cannot accurately reflect bacterial activity, and the instruments and equipment are extremely complicated, and the technical requirements for operation are particularly high and the detection throughput is low[ Reference 8]

The method based on Raman signal is also to judge the effect of antibiotics by detecting the metabolic activity of bacteria. This method is simple to operate and the measurement time is short, but there are still weak Raman signals, inaccurate detection of MIC for some antibiotics, and limited by sample purity, etc. Large background interference, low signal-to-noise ratio, poor compatibility with existing instruments (not as common as microplate readers, flow cytometers, etc.), relatively expensive prices, etc. [References 9, 10, 11]

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