Method for distinguishing fungi and bacteria
A technology for bacteria and fungi, applied in biochemical equipment and methods, microbial determination/inspection, fluorescence/phosphorescence, etc., can solve the problems of complicated preparation, high cost, etc., to reduce drug resistance, simple sample processing, and fast processing. Effect
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Example 1, PFP-NMe 3 + with PPV-NMe 3 + Synthesis
1. PFP-NMe 3 + Synthesis
1. Synthesis of 1,4-benzenediboronic acid neopentyl diester
2.2g of 1,4-phenylenediboronic acid, 50mL of toluene, 5.2g of neopentyl glycol and 0.5g of p-toluenesulfonic acid were added to a 100mL single-neck bottle, and refluxed for 24h; after the solvent was evaporated, silica gel column separation (eluent: dichloromethane) 4.8 g of product were obtained afterward. Characterization of the product: 1 H NMR (400MHz, CDCl 3 ): δ (ppm) 7.78 (s, 4H), 3.77 (s, 8H), 1.02 (s, 12H). Characterization data indicated that the product was 1,4-benzenediboronic acid neopentyl diester.
2. Synthesis of 2,7-dibromo-9,9-bis(6-iodohexyl)fluorene
Add 10 mL of dry tetrahydrofuran and 0.65 g of 2,7-dibromofluorene to a 50 mL single-neck flask, cool to -78°C at low temperature, add 2 mL of n-hexane containing butyllithium (the concentration of butyllithium is 2M), stir at low temperature for 1 h, add 0.67g 1,...
Example Embodiment
Example 2, PFP-NMe 3 + and PPV-NMe 3 + Application in detection of Escherichia coli [JM109]
1. Prepare the solution of pathogenic bacteria to be tested
Escherichia coli JM109 strain (Beijing Biomed Technology Development Co., Ltd.; product catalog number: CC0301) was added to 3 mL of LB medium, and cultured at 37° C. overnight. Take a certain volume of bacterial liquid, centrifuge at 4000 rpm for 5 min to remove the culture medium, wash three times with phosphate buffer solution (PBS, 10mM, pH=7.40), and finally adjust the bacterial liquid concentration in phosphate buffer solution to OD 600 = 1.0.
2. Direct observation under UV light
The PFP-NMe prepared in Example 1 3 + Dissolved in double distilled water to give PFP-NMe at a concentration of 1 mM 3 + solution. The PPV-NMe prepared in Example 1 3 + Dissolved in double distilled water to give PPV-NMe at a concentration of 1 mM 3 + solution.
Add 2 μL of PFP-NMe to 2 mL of 40% (volume percent) ethanol aqueou...
Example Embodiment
Example 3, PFP-NMe 3 + and PPV-NMe 3 + Application in the detection of Escherichia coli [M15]
1. Prepare the solution of pathogenic bacteria to be tested
The JM109 strain was replaced with Escherichia coli M15 strain (Beijing Bomed Technology Development Co., Ltd., CC0901), and other steps were the same as the first step in Example 2.
2. Direct observation under UV light
The JM109 bacterial solution was replaced by the M15 bacterial solution, and other steps were the same as the third step in Example 2.
Three parallel measurements were taken, and the average value was taken. The results are shown in Table 2.
Table 2 Fluorescence spectrum measurement results
The quenching efficiency of the M15 (bacteria) bacterial solution was 80.5%, which was greater than 50%, and the histogram was shown in Figure 2.
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