Method for simultaneously classifying and detecting carbapenemase, AmpC enzyme and extended-spectrum beta-lactamase

A carbapenemase, classification detection technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of high cost, inability to detect carbapenemase, IPM paper detection problems such as low detection rate, high detection rate, easy popularization and application, and low cost

Inactive Publication Date: 2016-10-26
明德松
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Problems solved by technology

[0003] In order to solve the above problems, the applicant published a multi-substrate synergistic-antagonistic method for simultaneous detection of extended-spectrum β-lactamase and AmpC β-lactamase (Zhang Guowei, Ming Desong, Wu Yibo. Multi-substrate synergistic-antagonistic method Simultaneous detection of extended-spectrum β-lactamase and AmpC β-lactamase [J]. Chinese Journal of Infection Control. 2003 (04)) This method sticks imipenem (IMP) antibiotic paper to the center of 9cm medium, ceftazidime (CAZ), ceftazidime / clavulanic acid, cefotaxime (CTX), cefotaxime / clavulanic acid, cefepime (FEP), and cefoxitin (FOX) are placed in sequence with the IPM as the center, and the The distance is 25mm, the distance between two adjacent discs is ≥24mm, the distance between cefotaxime / clavulanic acid and FEP is 20mm, and incubated at 35°C for 16-18h; the cost of this method is high, and it cannot be used for the detection of carbapenemase at the same time, and IPM The detection rate of paper chips is low, so it needs to be improved

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  • Method for simultaneously classifying and detecting carbapenemase, AmpC enzyme and extended-spectrum beta-lactamase

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specific Embodiment 1

[0037] Specific embodiment 1: with reference to figure 1As shown, the simultaneous classification detection method of carbapenemase, AmpC enzyme and extended-spectrum β-lactamase was applied to the detection of Acinetobacter isolated from blood samples of fracture wound infection, and the results of bacterial drug sensitivity showed that penicillins, The third-generation cephalosporins are highly drug-resistant. The bacteria to be tested in the blood sample were made into a suspension of 0.5 McGregor units and spread evenly on the surface of the M-H plate. The EDTA paper was placed in the center of the surface of the 9 cm plate, and the EDTA paper and ceftazidime ( CAZ) antibiotic disk, ceftazidime / clavulanic acid (CAZ / CLAV) antibiotic disk and ertapenem (ETP) antibiotic disk, piperacillin / tazobactam (TZP) antibiotic disk and cefoxitin ( The distance between FOX) antibiotic discs was adjusted to 15mm, and incubated overnight at 35°C. The results were as follows: the diameter o...

specific Embodiment 2

[0038] Specific embodiment 2: with reference to figure 1 As shown, the method of simultaneous classification and detection of carbapenemase, AmpC enzyme and extended-spectrum β-lactamase was applied to the detection of Acinetobacter isolated from blood samples of intracranial infection, and the bacterial drug sensitivity results showed that penicillins, third Alternative cephalosporins, ceftazidime (CAZ), imipenem (IMP) and meropenem (MER) are all highly drug-resistant, and the bacteria to be tested in the blood sample are made into a suspension of 0.5 McGregor units and spread evenly on the surface of the M-H plate. Paste ceftazidime (CAZ) antibiotic strips, ertapenem (ETP) antibiotic strips, ceftazidime / clavulanic acid (CAZ / CLAV) antibiotic strips, imipenem (IMP) antibiotic strips, Racicillin / tazobactam (TZP) antibiotic disc and cefoxitin (FOX) antibiotic disc, adjust the distance between EDTA disc and ceftazidime (CAZ) antibiotic disc to 15mm, ceftazidime / clavulanic acid (C...

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Abstract

The invention relates to the technical field of clinical microorganism bacterial detection, in particular to a method for simultaneously classifying and detecting carbapenemase, AmpC enzyme and extended-spectrum beta-lactamase. The method includes the steps that antibiotic test paper and EDTA test paper are selected for use, detected drug-resistant to-be-detected bacteria are prepared into a 0.5 McFarland unit bacterium suspension and evenly smeared to the surface of an M-H plate, the test paper containing EDTA is placed in the center of the surface of the M-H plate coated with the to-be-detected bacteria, ceftazidime (CAZ) antibiotic test paper, ertapenem (ETP) antibiotic test paper, piperacillin / tazobactam (TZP) antibiotic test paper, ceftazidime / clavulanic acid (CAZ / CLAV) antibiotic test paper, imipenem (IMP) antibiotic test paper, and cefoxitin (FOX) antibiotic test paper are attached around the EDTA test paper, gaps are reserved between the EDTA test paper and other antibiotic test paper and between every two pieces of antibiotic test paper, incubation is conducted for 16-18 h at 35 DEG C, and antibacterial zone diameters of the EDTA test paper and other antibiotic test paper are observed and recorded respectively. Cost is low, the detection rate is high, and application and popularization in a clinical laboratory are easy.

Description

technical field [0001] The invention relates to the technical field of clinical microbial bacterial detection, in particular to a method for simultaneously classifying and detecting carbapenemase, AmpC enzyme and extended-spectrum beta-lactamase. Background technique [0002] The β-lactamases associated with severe bacterial resistance are mainly carbapenemase, AmpC β-lactamase and extended-spectrum β-lactamase. Bacteria such as Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Klebsiella pneumoniae, Escherichia coli, etc. produced during the growth process can hydrolyze various β-lactam antibiotics, such as ertapenem ( ETP), meropenem (MER), ceftazidime (CAZ), cefoxitin (FOX) and other enzymes, some bacteria can produce multiple β-lactamases at the same time, and these enzyme-producing bacteria are resistant to the above antibiotics, So far, there is still a lack of simple and effective methods for the routine detection of enzyme-producing bacteria. [0003] In order ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34
CPCC12Q1/34
Inventor 明德松
Owner 明德松
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