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A screening chromogenic plate for carbapenem-resistant Enterobacteriaceae bacteria

A technology for antibiotics Enterobacteriaceae and carbapenems, which is applied in the field of screening color development plates for carbapenem-resistant Enterobacteriaceae bacteria. etc. to achieve good sensitivity and specificity, promote enzyme activity, and improve selectivity

Active Publication Date: 2022-05-27
浙江夸克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The bacteriological culture method needs to go through enrichment culture, selective separation, observation of morphological characteristics, physiological and biochemical reactions, serological identification and other processes, generally takes 4 to 7 days, the operation is cumbersome, time-consuming and labor-intensive, with low sensitivity and specificity, and Damaged and dead bacteria cannot be detected; immunological methods are prone to contamination, and have serious cross-reactions, many false positives, and low sensitivity; the biggest advantage of nucleic acid probe detection technology is its strong specificity, but there are also problems in probe detection technology. Certain problems, such as the sensitivity is not high enough; to detect a kind of bacteria, it is necessary to prepare a probe
The disk method is also a commonly used method for rapid detection of coliform bacteria; however, it is easily affected by the nature of the product to be tested, resulting in a decrease in accuracy

Method used

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  • A screening chromogenic plate for carbapenem-resistant Enterobacteriaceae bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] A preparation method of yeast extract, comprising the following steps:

[0054] (1) Take the yeast emulsion with an effective content of 6%, add 0.06g / L of citric acid and 0.15g / L of potassium chloride at 45°C, and autolyse at a constant temperature, and add 0.06g / L in a mass ratio of 1: 3:1 β-xylosidase, papain and glucanase (enzyme activities are 6000, 6000, 10000 U / g respectively), enzymatic hydrolysis at 35 ° C and pH value of 6.5; the enzymatic hydrolysis solution is obtained;

[0055] (2) Take the supernatant of the enzymatic hydrolysis solution and concentrate under reduced pressure to obtain a yeast extract.

[0056] The specific chromogenic substrate is 5-bromo-6-chloro-3-indole-β-glucoside with a concentration of 3.00g / L and 5-bromo-4-chloro-3-indole with a concentration of 1.00g / L - Beta galactoside.

[0057] The carbapenem antibiotic is a mixture of imipenem, meropenem and ertapenem;

[0058] Wherein, the effective concentration of imipenem is 10 mg / L; th...

Embodiment 2

[0067] (1) Take the yeast emulsion with an effective content of 10%, add 0.05g / L of citric acid and 0.2g / L of potassium chloride at 40°C, and autolyse at a constant temperature, and the mass ratio of adding 0.05g / L is 1: 2:2 β-xylosidase, papain and glucanase (enzymatic activities were 5000, 5000, 20000 U / g respectively), enzymatic hydrolysis at 30 ° C and pH value of 5; the enzymatic hydrolysis solution was obtained;

[0068] (2) Take the supernatant of the enzymatic hydrolysis solution and concentrate under reduced pressure to obtain a yeast extract.

[0069] The specific chromogenic substrate is 5-bromo-6-chloro-3-indole-β-glucoside with a concentration of 0.01g / L and 5-bromo-4-chloro-3-indole with a concentration of 5.00g / L - Beta galactoside.

[0070] The carbapenem antibiotic is a mixture of imipenem, meropenem and ertapenem;

[0071] Wherein, the effective concentration of imipenem is 0.1 mg / L; the effective concentration of meropenem is 50 mg / L; the effective concent...

Embodiment 3

[0080] (1) Take yeast emulsion with an effective content of 5%, add 0.1g / L of citric acid and 0.1g / L of potassium chloride at 60°C, and autolyse at a constant temperature, and add 0.1g / L in a mass ratio of 1: 3:1 β-xylosidase, papain and glucanase (enzyme activities are 6000, 5000, 8000 U / g respectively), enzymatic hydrolysis at 45 ° C and pH value of 7; an enzymatic hydrolysis solution is prepared;

[0081] (2) Take the supernatant of the enzymatic hydrolysis solution and concentrate under reduced pressure to obtain a yeast extract.

[0082] The specific chromogenic substrate is 5-bromo-6-chloro-3-indole-β-glucoside with a concentration of 5.00g / L and 5-bromo-4-chloro-3-indole with a concentration of 0.01g / L - Beta galactoside.

[0083] The carbapenem antibiotic is a mixture of imipenem, meropenem and ertapenem;

[0084] Wherein, the effective concentration of imipenem is 50 mg / L; the effective concentration of meropenem is 0.1 mg / L; the effective concentration of ertapenem...

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Abstract

The invention belongs to the technical field of bacteria detection, and in particular relates to a color-developing plate for screening Enterobacteriaceae bacteria resistant to carbapenem antibiotics. The chromogenic plate contains a specific chromogenic substrate and a selective medium; wherein, the selective medium includes the following components: yeast extract, salicin, gum arabic, sodium citrate, folic acid, vitamin B1, vitamin E. The chromogenic plate can be used for rapid screening of clinical carbapenem-resistant strains, and has a high accuracy rate.

Description

technical field [0001] The invention belongs to the technical field of bacterial detection, in particular to a screening color plate for carbapenem-resistant Enterobacteriaceae bacteria. Background technique [0002] Enterobacteriaceae are gram-negative bacilli, including Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, etc., which often exist in the upper respiratory tract and intestinal tract of the human body. infection. [0003] Imipenem and meropenem are commonly used carbapenem antibiotics in clinical practice. They have the characteristics of broad antibacterial spectrum, strong antibacterial activity, stability to β-lactamase, and low toxicity. One of the most important antibacterial drugs for infection was once considered to be a β-lactam antibacterial drug stable against ESBLs and AmpC enzymes. However, with the increase in clinical use of carbapenems, carbapenems Antibiotic-resistant bacteria are also on the rise. The emergence and rapid spread of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/18C12Q1/10
CPCC12Q1/18C12Q1/10C12Q1/045Y02A50/30
Inventor 章晓庆娄斌
Owner 浙江夸克生物科技有限公司
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