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Carbapenemase-producing strain rapid detection method and kit, and application of kit

A carbapenemase and kit technology, applied in the biological field, can solve the problems of long detection time and achieve the effect of simple operation, high accuracy and specificity

Inactive Publication Date: 2019-03-29
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, an obvious shortcoming of this method is that it takes 18 to 24 hours to get the test results, and the test time is long.
The Carba NP test and Carba NP direct test have higher technical requirements, and the scope of application is not as wide as that of CIM, requiring special reagents and professional operation

Method used

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  • Carbapenemase-producing strain rapid detection method and kit, and application of kit
  • Carbapenemase-producing strain rapid detection method and kit, and application of kit
  • Carbapenemase-producing strain rapid detection method and kit, and application of kit

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Experimental program
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Effect test

Embodiment 1

[0053] The construction of embodiment 1 detection method

[0054] figure 1 It is a schematic flowchart of the method for rapidly detecting carbapenemase-producing bacteria of the present invention. The method includes the following steps:

[0055] Step S101: Place the imipenem drug-sensitive disc in a centrifuge tube containing sterile water, and vortex the centrifuge tube.

[0056] Imipenem drug-sensitive paper is a commercialized drug-sensitive paper. Place the imipenem drug-sensitive paper in a centrifuge tube containing sterile water for vortexing, mainly to evenly diffuse the drug in the paper in sterile water.

[0057] In this embodiment, the step of placing the imipenem drug-sensitive paper disc in a centrifuge tube containing sterile water, and vortexing the centrifuge tube includes: placing a 10 μg imipenem drug-sensitive disc in Place in a 2 ml centrifuge tube containing 500 µl sterile water and vortex the tube for 10-20 seconds. Place the imipenem drug-sensitiv...

Embodiment 2

[0082] (1) Set up a negative control group ( figure 2 negative sign "-" position), a positive control group ( figure 2 The positive sign "+" position), an experimental group ( figure 2 No. 2 position in middle), wherein the negative control group is a standard strain of Escherichia coli ATCC25922 (purchased from China Microorganism Culture Collection Center) which has been identified by PCR without enzyme gene, and the positive control group is a strain of Klebsiella pneumoniae ATCC BAA- 1705 (purchased from China Microorganism Culture Collection), and the experimental group was a strain of Klebsiella pneumoniae ATCCBAA2146 (purchased from China Microorganism Culture Collection).

[0083] After the CIM method (method refers to the literature Zwaluw K V D, Haan A D, Pluister G N, et al. The Carbapenem Inactivation Method (CIM), a Simple and Low-Cost Alternative for the Carba NP Test to Assess Phenotypic Carbapenemase Activity in Gram-NegativeRods[J]. Plos One,2015,10(3):e0...

Embodiment 3

[0101] (1) Set up a negative control group ( image 3 negative sign "-" position), a positive control group ( image 3 The positive sign "+" position), an experimental group ( image 3 No. 3 position in middle), wherein the negative control group is an Escherichia coli ATCC25922 (purchased from China Microorganism Culture Collection Center) that has no enzyme-producing gene identified by PCR, and the positive control group is a strain of Klebsiella pneumoniae ATCCBAA-1705 (purchased The experimental group was a Klebsiella pneumoniae ATCC BAA-1706 (purchased from the China Microorganism Collection).

[0102] Draw experimental result through CIM method (with embodiment 2): above-mentioned negative control group is the bacterium that does not produce carbapenemase, and positive control group is the bacterium that produces carbapenemase, and experimental group is the bacterium that does not produce carbapenemase Enzyme bacteria.

[0103] (2) Place a piece of imipenem drug-sensi...

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Abstract

The invention provides a carbapenemase-producing strain rapid detection method and kit, and an application of the kit. The rapid detection method includes the steps: carrying out common incubation ofimipenem with a to-be-tested bacterial lawn to prepare an MH agar culture medium containing bromcresol purple and acid-producing bacteria, pasting imipenem paper after incubation with the to-be-testedbacterial lawn on the MH agar culture medium containing the bromocresol purple and the acid-producing bacteria, incubating for 3.5-4.5 h, and determining whether the to-be-tested strain is a carbapenemase-producing strain directly through a fact whether bromocresol purple is discolored. The detection method has the advantages of simple operation, fast analysis, wide application range, high accuracy and specificity, and easy interpretation of the results. At the same time, the invention also provides the detection kit established based on the rapid detection method; the kit has important significance for early discovery of the carbapenemase-producing strain and adopting of effective measures, nosocomial infection control and clinical epidemiological detection, and has good application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for rapidly detecting carbapenemase-producing strains, a kit and an application thereof. Background technique [0002] Carbapenem antibiotics have significant therapeutic effects in the treatment of infections caused by most Gram-positive and Gram-negative bacteria, so they have a very important role in clinical practice. However, with the continuous evolution of bacteria, some bacteria can produce carbapenemase, that is, with the emergence of carbapenemase-producing bacteria, many cases of clinical failure of carbapenem drugs appear. [0003] At present, the carbapenem inactivation method (CIM) is a commonly used method for clinical detection of carbapenemase-producing strains. It is widely used because of its low price and simple operation. However, an obvious shortcoming of this method is that it takes 18 to 24 hours to obtain the detection result, and the detection time...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/125C12R1/07
CPCC12Q1/04G01N2333/32
Inventor 孙坚崔泽华胡佳玲贾玲唐甜刘雅红廖晓萍卢思雅
Owner SOUTH CHINA AGRI UNIV
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