Detection reagent and method for detecting enzyme type of carbapenemase

A carbapenemase enzyme and detection reagent technology, applied in the field of carbapenemase enzyme type detection, can solve the problems of high mortality rate of patients, poor antibiotic treatment effect, etc., and achieves simple operation, easy-to-read results and low cost low effect

Pending Publication Date: 2022-01-28
中国人民解放军西部战区总医院
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Problems solved by technology

[0002] Epidemiology in recent years suggests that with the widespread use of broad-spectrum antibiotics and carbapenem antibiotics in disease infections, the incidence of Carbapenem-resistant Enterobacteriaceae (CRE) infections has increased year by year, and many A study suggests that if CRE infection occurs, the effect of traditional antibiotic treatment is poor, and the mortality rate of patients is high

Method used

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  • Detection reagent and method for detecting enzyme type of carbapenemase
  • Detection reagent and method for detecting enzyme type of carbapenemase
  • Detection reagent and method for detecting enzyme type of carbapenemase

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Embodiment 1

[0035] The embodiment of the present invention provides a detection reagent for detecting the enzyme type of carbapenemase, and its configuration method is as follows:

[0036] Configure the first solution, 0.1mol / L EDTA, and use 0.5mol / L EDTA solution to configure 0.1mol / L: Specifically, take 0.5mol / L EDTA200ul and add 800ul sterilized water to obtain 0.5M EDTA in one milliliter .

[0037] Prepare the second solution: 50mg / ml 3-aminophenylboronic acid: Mix 25mg of triaminophenylboronic acid, 250ml of distilled water and 250ml of dimethyl sulfoxide to obtain 500ml of 50mg / ml 3-aminophenylboronic acid. The obtained first liquid and second liquid are diluents for detecting the enzyme type of carbapenemase. Subpackage, that is.

Embodiment 2

[0039] This embodiment provides a method for detecting the enzyme type of carbapenemase, including: using the detection reagent in Example 1 to detect, specifically:

[0040] (1): Adjust the concentration of the CRE strain to be tested to 0.5 McFarland, dip the prepared bacterial solution with a sterile cotton swab, squeeze off the excess liquid on the test tube wall, and spread it on the upper surface of the M-H plate, rotating each time The flat plate is 60 degrees, and it is coated three times in total, and rubbed around the periphery twice to make the entire flat plate evenly coated.

[0041] (2): Place the plate for 3-5 minutes, use sterile tweezers to take the drug-sensitive paper, and paste 4 pieces of imipenem paper on the MH plate coated with the CRE strain to be tested, and the paper is evenly distributed. The distance between the sheets shall not be less than 24mm, and the distance between the center of the sheet and the edge of the plate shall not be less than 15mm...

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Abstract

The invention relates to the technical field of detection of the enzyme type of carbapenemase, in particular to a detection reagent and method for detecting the enzyme type of the carbapenemase. The raw materials of the detection reagent comprise an ethylenediamine tetraacetic acid compound and triaminophenylboronic acid, the concentration of a solution containing the ethylenediamine tetraacetic acid compound is 0.8-1.2 mol / L, and the concentration of a solution containing the triaminophenylboronic acid is 40-60 mg / ml. The detection reagent not only can realize typing of the carbapenemase, but also is simple to operate, easy in result reading and low in cost; and the detection reagent is very suitable for clinical carbapenemase phenotype detection.

Description

technical field [0001] The invention relates to the technical field of carbapenemase enzyme type detection, in particular to a detection reagent and method for detecting carbapenemase enzyme type. Background technique [0002] Epidemiology in recent years suggests that with the widespread use of broad-spectrum antibiotics and carbapenem antibiotics in disease infections, the incidence of Carbapenem-resistant Enterobacteriaceae (CRE) infections has increased year by year, and many This study suggests that if CRE infection occurs, the effect of traditional antibiotic treatment is poor, and the mortality rate of patients is high. Carbapenemase refers to a large class of β-lactamases that can hydrolyze carbapenem antibiotics, and can be divided into class A, class B and class D enzymes. Class A and Class D are serinases, and Class B are metalloenzymes. The class A KPC serine enzyme is the most important drug resistance mechanism of Enterobacteriaceae bacteria in recent years, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34
CPCC12Q1/34G01N2333/986
Inventor 汪璐曲远青刘媛蒲晓凤刘璐
Owner 中国人民解放军西部战区总医院
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