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Method for microbial antibiotic resistance detection

A technology of microorganisms and resistance, applied in the fields of biochemical equipment and methods, determination/inspection of microorganisms, etc.

Inactive Publication Date: 2005-07-27
EPPENDORF AG
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0009] Solve this problem by providing a method for detecting the presence or absence of microorganisms exhibiting resistance to β-lactam antibiotics in a biological sample and simultaneously determining the β-lactam resistance genotype

Method used

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  • Method for microbial antibiotic resistance detection
  • Method for microbial antibiotic resistance detection
  • Method for microbial antibiotic resistance detection

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Embodiment 1

[0162] design microarray

[0163] 1. Oligonucleotide Array Construction

[0164] The oligonucleotide array consisted of 168 oligonucleotide capture probes with varying lengths from 17 to 27 bases. Capture probe and control probe sequences are given in Table 2 (above), probes with a 15 thymidine spacer are marked with an asterisk. The array layout is shown in Figure 1.

[0165] Oligonucleotide microarrays were formed to determine TEM beta-lactamase genotypes. For 41 SNP (Single Nucleotide Polymorphism) sites (ESBL, IRT or both), design oligonucleotide probes of various lengths (17-27 bases) (Table 2). These probes cover most of the amino acid substitution sites currently known in the art, except for amino acids 2, 3 and 285, which are common at the end of TEM beta-lactamase.

[0166] Probes are designed so that the SNP is at an essentially central base in the probe sequence to maximize discrimination between perfect matches / mismatches during hybridization. For each SNP,...

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Abstract

The present invention relates to methods for fast detecting the fastness of a micro-organism beta-lactam utilizing micro-arrays, and / or methods for fast detecting coding micro-organism beta-lactam enzymatic nucleotideacid sequence, and the detecting reagent box thereof.

Description

technical field [0001] The invention relates to a method for rapidly detecting the resistance of microorganisms to antibiotics by using a microarray and simultaneously determining the genotype of the resistance genotype, and / or a kit for the method. Background technique [0002] In the past, the continued use of antibiotics, especially in clinical settings, has increased the selective pressure on bacteria, eventually resulting in the development of many microorganisms that exhibit resistance to the agents used. Bacteria produce enzymes called β-lactamases for one major class of antibiotics that rely on the β-lactam ring system. This enzyme is capable of degrading such reagents and is already ubiquitous in different microbial species. [0003] To date, 340 different β-lactamase sequences have been reported. TEM β-lactamase is a serine β-lactamase (ie, OXA, SHV) found in many different species of Enterobacteriaceae, Pseudomonas aeruginosa, Haemophilus influenzae, and Neisser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/689C12Q1/6837C12Q1/6876C12Q2600/156C12Q2565/519C12Q2525/179
Inventor 韦雷娜·容桑托西·埃扎奇米洛拉德·苏沙科尔内留斯·克纳比罗尔夫·施密特齐尔·T·巴赫曼
Owner EPPENDORF AG