Bacterial or fungal activity rapid detection method

A detection method, a bacterial technology, applied in the biological field, can solve the problems of complex operation of MIPs, unstudied MIPs to measure the activity of bacteria or fungi, etc., and achieve the effect of improving the effect of imprinting

Active Publication Date: 2020-04-14
武汉南嘉木实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above studies have shown that bacterial MIPs can specifically bind and detect target bacteria, but none of the above work has studied whether MIPs can measure the activity of bacteria or fungi, and the operation of the above-mentioned MIPs for detecting bacteria or fungi is relatively complicated

Method used

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  • Bacterial or fungal activity rapid detection method
  • Bacterial or fungal activity rapid detection method
  • Bacterial or fungal activity rapid detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Preparation of fluorescent molecularly imprinted polymer I for Escherichia coli

[0053] The method for preparing the fluorescent molecularly imprinted polymer I of Escherichia coli comprises the steps of:

[0054] 1) Preparation of reagents:

[0055] Dopamine solution Ⅰ: ammonium persulfate and dopamine are dissolved in 10mmol / L Tris-HCl buffer solution (pH=8 of Tris-HCl buffer solution) at a molar ratio of 2:1;

[0056] Dopamine solution II: dopamine dissolved in 10mmol / L Tris-HCl buffer solution (Tris-HCl buffer solution pH=8);

[0057] Dopamine / dansyl dopamine mixed solution: dopamine: dansyl dopamine: ammonium persulfate molar ratio=4:1:1 molar ratio dissolved in 10mmol / L Tris-HCl buffer solution (Tris-HCl buffer solution pH=8) ;

[0058] Carrier material: 96-well microtiter plate is used as the carrier material;

[0059] Target cells: 10 4 ~10 6 CFU / mL Escherichia coli liquid, E. coli285 is used for Escherichia coli;

[0060] Eluent: deionized wat...

Embodiment 2

[0067] Example 2 Preparation of fluorescent molecularly imprinted polymer II for Escherichia coli

[0068] Compared with Example 2, the preparation method of Example 2 differs in that Example 2 does not carry out step 2) in Example 1, that is, Example 2 does not use the polydopamine-modified dopamine-carrier complex as a carrier, and It is to directly polymerize dopamine, dansyl dopamine, and Escherichia coli on the carrier material. The specific process is as follows:

[0069] 1) Preparation of Escherichia coli-dansyl dopamine / dopamine-carrier complex: add dopamine / dansyl dopamine mixed solution and target cells (E. 37°C, the rotation speed is 150rpm) shake slowly for 72h, then pour out the liquid in the hole, and obtain the Escherichia coli-dansyl dopamine / dopamine-carrier complex;

[0070] 2) Elution of target cells: wash the Escherichia coli-dansyldopamine / dopamine-carrier complex repeatedly with an eluent, and elute the Escherichia coli in the Escherichia coli-dansyldop...

Embodiment 3

[0071] Example 3 Preparation of fluorescent molecularly imprinted polymer III for Escherichia coli

[0072] The preparation method of the fluorescent molecularly imprinted polymer III of Escherichia coli is the same as that of Example 1, except that in this example, the dopamine solution I in step 2) of Example 1 is changed to dopamine solution II, and the dopamine solution I and dopamine solution The difference of II is whether ammonium persulfate is added or not. The method for preparing the fluorescent molecularly imprinted polymer III of Escherichia coli in this example will not be repeated here.

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Abstract

The invention relates to the biotechnology field, in particular to an activity fast detection method of bacteria or fungi. The activity fast detection method of bacteria or fungi comprises the steps that (1) firstly, a dopamine / dabsyl-L-proline dopamine mixed solution is prepared; then, the dopamine / dabsyl-L-proline dopamine mixed solution, target cells and carriers are uniformly mixed; polymerization reaction is performed at the surfaces of the carriers to obtain a target cell-dabsyl-L-proline dopamine / dopamine-carrier compound body; finally, the target cells are eluted from the compound body by an eluting agent; a fluorescent molecular imprinting polymer of the target cells is obtained; (2) after samples are added into the fluorescent molecular imprinting polymer, the fluorescence value is immediately measured as I0; after the bacteria or fungi in the samples are completely combined with the fluorescent molecular imprinting polymer, the fluorescence value is measured as I; I0 / I-1 expresses the fluorescence change level; the content of target bacteria or target fungi in the samples is determined through the fluorescence change level. The activity of the bacteria or fungi in the samples can be fast and accurately detected by the method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid detection method for the activity of bacteria or fungi. Background technique [0002] Bacteria and fungi widely exist in nature and the human body, and pathogenic bacteria and fungi can seriously endanger the health of living organisms. According to statistics, about one-third of deaths worldwide are related to bacterial infectious diseases. Pathogenic bacteria infect the body to cause disease, usually need: ① invade and colonize the body; ② proliferate in the host body and spread to other parts; ③ evade the body's defense mechanism; ④ release toxins or trigger hypersensitivity reactions. The invasion of active pathogenic bacteria into the body is the first step of bacterial infection and the first condition for pathogenic bacteria to cause disease in the body. Therefore, rapid, accurate and sensitive detection methods for bacterial activity are very important to reduce the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 吕斌刘燕婕李宁叶磊吴中乔邓耘项阳
Owner 武汉南嘉木实业有限公司
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