Method for quantitatively analyzing escherichia coli drug resistance based on golden shell-up-conversion chiral dipolymer
A technology for quantitative analysis of Escherichia coli, applied in the field of materials chemistry, to achieve the effect of short time, high sensitivity and low detection limit
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Embodiment 1
[0035] Example 1 Preparation of gold shell-up-conversion chiral dimer
[0036] All glass instruments were soaked in aqua regia, rinsed with double distilled water, and dried for later use. The water used in the experiment is 18.2MΩ Milli-Q ultrapure water.
[0037] (1) Synthesis of gold nanoparticles: Gold nanoparticles with a diameter of 10±2 nm were synthesized by reducing chloroauric acid with tannic acid, and 0.1 mL of 1% tannic acid aqueous solution and 80 mL of 0.0125% chloroauric acid aqueous solution were heated to 60°C, then quickly add the tannic acid solution into the chloroauric acid solution under vigorous stirring. After reacting for 2 h, when the color changed, the solution was cooled to room temperature, and then the solution was stored at 4°C for later use.
[0038] (2) Preparation of gold-shell nanoparticles: First, centrifuge the above-prepared gold nanoparticles at 13,000 rpm for 10 min, concentrate the precipitate 5 times and resuspend in phosphate buffe...
Embodiment 2
[0043] Example 2 Detection of gold shell-up-converting chiral dimer drug resistance to Escherichia coli
[0044] (1) Bacterial sources and culture conditions: Polymyxin B-resistant E. coli strains (E15017, resistance level 24 mg / mL), polymyxin B-sensitive E. coli strains (MG1655, drug resistance level 4 mg / mL) were obtained from Provided by Zhejiang University. Polymyxin B-resistant Escherichia coli strains (EYAC08-25-1, resistance level 20mg / mL and GZP11-11, resistance level 16mg / mL) were provided by China Agricultural University. Polymyxin B-resistant Escherichia coli strain (JN-PBR-1, resistance level 8 mg / mL) was obtained from Jiangnan University. A single colony on a solid agar plate was inoculated into 10 mL of LB medium and cultured overnight at 37 °C with constant shaking at 200 rpm. Bacteria in exponential growth phase were obtained, OD 600 is 0.7 (5×10 7 CFU / mL). Then, the bacteria were diluted to a final concentration of 1 × 10 by PBS buffer (pH 7.4) 3 CFU / m...
Embodiment 3
[0046] Example 3 Characterization of gold shell-upconversion chiral dimer for drug resistance detection of Escherichia coli: the formation process of the chiral dimer was characterized by electron microscopy; the fluorescence signal and CD signal were characterized for the detection of bacterial drug resistance, and A standard curve was established separately. like figure 2 As shown in b and 3b, the degree of drug resistance of E. coli has a good linear relationship with the ratio of the number of bacteria to the CD signal and the ratio of the number of bacteria to the fluorescence intensity, respectively. It shows that this method can be used to detect the degree of drug resistance of Escherichia coli.
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