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Fluorescent probe and method for rapidly detecting staphylococcus aureus by using same

A technology of fluorescent probes and staphylococci, which is applied in the field of environmental sanitation and renewable resource chemical biology, can solve the problems of high technical operation requirements, expensive instruments, unfavorable popularization, etc., and achieve simple process, simple operation, stable and controllable fluorescence The effect of the characteristic

Inactive Publication Date: 2011-05-11
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The use of some instruments can effectively reduce the detection time of Staphylococcus aureus to achieve rapid and specific identification of Staphylococcus aureus, such as enzyme-linked immunoassay (PCR), RNA probe, and flight mass spectrometry, but the required instruments are expensive. High technical operation requirements are not conducive to popularization

Method used

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  • Fluorescent probe and method for rapidly detecting staphylococcus aureus by using same
  • Fluorescent probe and method for rapidly detecting staphylococcus aureus by using same
  • Fluorescent probe and method for rapidly detecting staphylococcus aureus by using same

Examples

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Embodiment 1

[0027] Carboxymethyl chitosan was dissolved in distilled water at room temperature to configure a concentration of 5mg / ml solution, and 5×10 -3 mol / l cadmium acetate solution, and stirred for 8h to obtain carboxymethyl chitosan cadmium complex. The complex solution was placed in a three-necked flask, protected by nitrogen gas, and quickly added a freshly configured aqueous sodium sulfide solution under high-speed stirring at 2000r / m so that the molar ratio of Cd:S was 1:2, and then continued to mix under nitrogen protection. The system was heated to 80-100 degrees and refluxed for 2 hours to obtain a carboxymethyl chitosan / CdS quantum dot composite solution. The above product was dialyzed in pH=7.5 phosphate buffer for 2 days, purified by gel filtration chromatography (column filler was Sephedex G100), and concentrated to obtain a purified aqueous solution of quantum dots. Dissolve carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in distilled water to make a 500mM solution, ...

Embodiment 2

[0029] Dissolve carboxymethyl chitosan in distilled water at room temperature to configure a concentration of 5mg / ml solution, and add 1×10 -2mol / l cadmium acetate solution, and stirred for 8h to obtain carboxymethyl chitosan cadmium complex. Put the complex solution in a three-necked flask, protect it with nitrogen gas, and quickly add a freshly configured sodium selenide aqueous solution under high-speed stirring at 2000r / m, so that the molar ratio of Cd:Se is 1:1, and then continue under nitrogen protection. The temperature of the system was raised to 80-100 degrees and refluxed for 2 hours to obtain a carboxymethyl chitosan / CdSe quantum dot composite solution. The above product was dialyzed in pH=7.5 phosphate buffer for 2 days, purified by gel filtration chromatography (column filler was Sephedex G100), and concentrated to obtain a purified aqueous solution of quantum dots. Dissolve carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in distilled water to make a 2 μM solut...

Embodiment 3

[0031] Carboxymethyl chitosan was dissolved in distilled water at room temperature to configure a concentration of 5mg / ml solution, and 5×10 -3 mol / l zinc acetate solution, and stirred for 8h to obtain carboxymethyl chitosan zinc complex. The complex solution is placed in a three-necked flask, protected by nitrogen, and is rapidly added with a freshly prepared sodium sulfide aqueous solution under high-speed stirring at 2500r / m so that the molar ratio of Zn:S is 1:1, and then continue to be mixed under nitrogen protection. The system was heated to 80-100 degrees and refluxed for 2 hours to obtain a carboxymethyl chitosan / ZnS quantum dot composite solution. The above product was dialyzed in pH=7.5 phosphate buffer for 2 days, purified by gel filtration chromatography (column filler was Sephedex G100), and concentrated to obtain a purified aqueous solution of quantum dots. Dissolve carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in distilled water to make a 2 μM solution, and...

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Abstract

The invention discloses a fluorescent probe, which is prepared by the following method. The method comprises the steps of: firstly, purifying fluorescence quantum dots synthesized in a carboxymethyl chitosan template, and connecting the fluorescence quantum dots with IgG (Immunoglobulin G) antibodies of a human by coupling effect of carbodiimide and N-hydroxysuccinimide eater; and then separating and purifying by using a sephadex column to obtain the fluorescent probe of the fluorescence quantum dots synthesized by immune-modified carboxymethyl chitosan. The invention further discloses a method for detecting staphylococcus aureus by using the fluorescent probe, comprising the detailed steps of: culturing the fluorescent probe and bacteria to be detected for 30 minutes, removing the fluorescent probe with nonspecific absorption by a plurality of times of centrifugal washing, finally, suspending bacteria marked by the fluorescent probe in a phosphate buffer solution, determining the fluorescence intensity and absorption intensity of bacterial suspension by adopting fluorescence spectrum and ultraviolet and visible absorption spectrum, and realizing quantitative determination of the staphylococcus aureus according to linear relation of the fluorescence intensity and the concentration of the bacterial suspension. The method disclosed by the invention is clean, simple and rapid and is suitable for being applied in rapid detection under emergency.

Description

technical field [0001] The invention relates to a method for specific rapid detection and identification of Staphylococcus aureus by using immunomodified carboxymethyl chitosan / quantum dots, which belongs to the field of chemical biology of renewable resources and the field of environmental sanitation. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus) is a common human pathogenic bacteria, easy to multiply in the human nasal cavity, skin and mucous membranes, causing various suppurative infections, such as phlebitis, osteomyelitis, endocarditis, secret Urinary tract infection, medical device, surgical infection, and food poisoning and toxic shock. The conventional detection and identification method of Staphylococcus aureus is mainly based on bacterial culture and colony counting, which takes about a week to complete. In the face of food safety and environmental safety emergencies, it is often required to complete the identification and detection o...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N21/64C09K11/89C09K11/88C09K11/56
Inventor 王小慧李栋孙润仓
Owner SOUTH CHINA UNIV OF TECH
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