Method for detecting drug resistance genes of soil microorganisms

A technology of soil microorganisms and detection methods, applied in the field of detection of soil microbial drug resistance genes

Active Publication Date: 2016-05-18
JIANGHAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the problem that the method for detecting drug resistance genes in the prior art needs to be improved, the embodiment of the present invention provides a method for detecting drug resistance genes of soil microorganisms

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  • Method for detecting drug resistance genes of soil microorganisms
  • Method for detecting drug resistance genes of soil microorganisms
  • Method for detecting drug resistance genes of soil microorganisms

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Experimental program
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Embodiment

[0030] Antimicrobial Resistance Gene Detection of Soil Microorganisms

[0031] Soil microorganisms come from the natural environment, and the drug resistance genes reflect the release of resistant drugs in the environment to a certain extent. The sample to be tested in this embodiment is soil, which is taken from a paddy field in Zhuankou Development Zone, Wuhan, Hubei. This embodiment includes the following steps:

[0032] Step 1. Determine the drug resistance gene of the microorganism to be detected in the sample to be tested, the endogenous standard gene in the sample to be tested and the exogenous standard gene of the sample to be tested. The specific methods are as follows:

[0033] Among them, there is at least one drug resistance gene, at least one endogenous standard gene, and at least one exogenous standard gene. The endogenous standard gene is the gene in the microorganism of the sample to be tested. An exogenous standard gene is not present in an organism of know...

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Abstract

The invention discloses a method for detecting drug resistance genes of soil microorganisms. The method comprises the following steps: determining drug resistance genes, standard endogenous genes and standard exogenous genes; preparing multiple amplification primers; adding exogenous nucleic acid into a to-be-tested sample, so as to obtain a mixed sample, and extracting a genome; adding the standard exogenous genes into the genome, so as to obtain mixed nucleic acid; amplifying mixed nucleic acid, constructing a high-throughput sequencing library by virtue of amplification products, and carrying out high-throughput sequencing, so as to obtain a sequencing segment group; analyzing the sequencing segment group, and judging whether the experiment is successful or not according to the number of obtained sequencing segments of the standard exogenous genes and the standard endogenous genes; if the experiment is successful, calculating the content of the resistance genes; and judging whether the to-be-tested sample contains the resistance genes according to the content of the resistance genes. By virtue of the method, a number of arbitrary drug resistance genes, which are expected to be detected, in arbitrary microorganism can be quantitatively detected once, furthermore, the microorganisms do not need to be cultured and purified during the detection, so that the detection speed is high, and the detection result is accurate and reliable.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting soil microbial drug resistance genes. Background technique [0002] Antibiotics are widely used in the prevention and treatment of animal and plant diseases, but the abuse of antibiotics will lead to the evolution of resistance genes in microorganisms. Monitoring microbial resistance genes in soil can reflect the use of antibiotics in the environment, and at the same time, can provide evidence for the effective use of antibiotics. [0003] Existing methods for detecting drug-resistant genes include: estimating the pathogenic bacteria of the drug-resistant gene that needs to be detected on the sample to be tested, isolating and purifying the pathogenic bacteria, and amplifying by PCR (Polymease Chain Reaction, polymerase chain reaction) primers For the drug resistance gene, electrophoresis or real-time quantitative PCR method is used to determine whether there ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2545/101C12Q2545/113C12Q2537/143
Inventor 李甜甜章伟雄彭海
Owner JIANGHAN UNIVERSITY
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