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Raman spectroscopy-heavy water isotope labeling based rapid detection method for drug susceptibility of drug-resistant bacteria and judgment method for rational drug use

A technology of isotope labeling and Raman spectroscopy, which is applied in the field of rapid detection of drug-resistant bacteria susceptibility and judgment of rational drug use, can solve the problems of lack of rapid detection technology of drug-resistant bacteria susceptibility, to curb the spread of bacterial resistance, analyze Simple, easy-to-operate effect

Active Publication Date: 2018-07-10
INST OF URBAN ENVIRONMENT CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As shown in the review, there is still a lack of rapid, easy-to-operate, and clinically applicable rapid detection technology for drug-resistant bacteria susceptibility

Method used

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  • Raman spectroscopy-heavy water isotope labeling based rapid detection method for drug susceptibility of drug-resistant bacteria and judgment method for rational drug use
  • Raman spectroscopy-heavy water isotope labeling based rapid detection method for drug susceptibility of drug-resistant bacteria and judgment method for rational drug use
  • Raman spectroscopy-heavy water isotope labeling based rapid detection method for drug susceptibility of drug-resistant bacteria and judgment method for rational drug use

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Example 1: Raman-heavy water isotope labeling method to quickly distinguish antibiotic-resistant and sensitive Escherichia coli

[0044] attached figure 1 The detection process of the Raman-heavy water isotope labeling method is given, including:

[0045] Add 200 μL of 10×~60×MIC antibiotics and 30%-100% (vol / vol) heavy water D in a 96-well plate 2 O (99.9atom% D; Sigma-Aldrich) LB broth, 200 μL without antibiotics but with heavy water D 2 O's LB medium was used as a control. Inoculate 5uL overnight cultured sensitive and drug-resistant bacteria in the culture medium. After incubating for 0, 0.5, 1, 1.5, and 2 hours at different times, the bacterial solution was taken out and washed by centrifugation with deionized water or 70% ethanol. The conditions were: 5000 rpm, 3 min each time, twice in total. Drop 2 μL of the cleaned bacterial solution on aluminum foil paper, dry it in the air, and perform Raman detection. Raman detection can be performed at the single-cell ...

Embodiment 2

[0047] Example 2: Raman-heavy water isotope labeling method to quickly detect the drug susceptibility of Escherichia coli to different antibiotics and guide rational drug use

[0048] Here, four different types of drug-resistant E. coli are used as examples, including ampicillin-resistant E. coli Escherichiacoli Amp R , Kanamycin-resistant Escherichia coli Kan R , Chloramphenicol-resistant Escherichiacoli Chl R , and the multidrug-resistant Escherichia coliT that is resistant to all three antibiotics R . The drug susceptibility of the above four bacteria has been tested by using the standard zone of inhibition test. The test results show that the sensitive Escherichia coli DH5α is sensitive to the three antibiotics, and Amp R Resistant to ampicillin, sensitive to kanamycin and chloramphenicol, Chl R Resistant to chloramphenicol, sensitive to ampicillin and kanamycin, Kan R Resistant to kanamycin, sensitive to ampicillin and chloramphenicol, T R Resistant to all three ant...

Embodiment 3

[0050] Example 3: Rapid detection of drug susceptibility of other pathogenic bacteria to different antibiotics by Raman-heavy water isotope labeling method

[0051] Three other pathogenic bacteria are used here as examples, including Salmonella enterica, Shigella flexneri, and Proteus vulgaris. In order to ensure the effectiveness of antibiotics and the accuracy of the discrimination results, Raman detection was also performed on a strain of quality control bacteria Escherichia coli 25922. Shigella flexneri and Proteus vulgaris were extracted from fresh chicken manure. The experimental procedure is basically the same as in Example 2, except that the incubation time is 1 hour. The results are attached Figure 4 as shown, Figure 4 It is above-mentioned four kinds of bacteria in 100% (vol / vol) heavy water D without antibiotics (control), and ampicillin, chloramphenicol, kanamycin, meropenem containing 10×MIC concentration 2 Relative C-D peak intensity ratio obtained by Raman...

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Abstract

The invention discloses a Raman spectroscopy-heavy water isotope labeling based rapid detection method for the drug susceptibility of drug-resistant bacteria and a judgment method for rational drug use. Directed at the disadvantage of long time in traditional drug susceptibility detection, the invention utilizes the principle that drug-resistant bacteria and sensitive bacteria have different activities under the action of antibiotics and accordingly C-D Raman peaks have different intensities, and realizes rapid detection of drug susceptibility within 0.5-1h. A to-be-detected object is incubated in a culture solution containing heavy water, wherein the group containing antibiotics is adopted as the treatment group, and the group without antibiotics is adopted as the control group, the incubated to-be-detected object is subjected to centrifugal cleaning, then Raman detection is carried out, the C-D / (C-D+C-H) values of the treatment group and the control group are calculated respectively,and the ratio of the treatment group and the control group is taken as the judgment standard, if the value is smaller than or equal to 0.75, the detected object can be susceptible to antibiotics, andif the value is greater than 0.75, the detected object can be resistant to the antibiotics. The method is simple, is easy for operation and analysis, and is suitable for different antibiotics and different bacteria, thus having clinical application prospects.

Description

technical field [0001] The invention relates to the technical field of biochemical detection and analysis, in particular to a method for rapid detection of drug susceptibility of drug-resistant bacteria based on Raman spectrum-heavy water isotope labeling and a method for judging rational drug use. Background technique [0002] The invention of antibiotics is one of the most important advances in the history of medicine. The use of antibiotics has effectively controlled bacterial infections and saved countless lives. The medical and health field accounts for half of the use of antibiotics (the other half is the aquaculture industry). However, the irrational use of antibiotics is very prominent. With the extensive use of antibiotics, the problem of bacterial drug resistance is becoming more and more serious, resulting in bacterial infections that cannot be effectively cured, and is showing an aggravating trend worldwide. Different countries, including developed countries (UK...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/65
Inventor 崔丽杨凯朱永官
Owner INST OF URBAN ENVIRONMENT CHINESE ACAD OF SCI
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