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A detection method for microbial drug resistance genes for non-disease diagnosis and treatment purposes

A detection method and drug resistance technology, applied in the biological field, to achieve the effect of strong versatility

Active Publication Date: 2019-11-15
JIANGHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 microbial drug resistance genes for non-disease diagnosis and treatment purposes

Method used

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  • A detection method for microbial drug resistance genes for non-disease diagnosis and treatment purposes
  • A detection method for microbial drug resistance genes for non-disease diagnosis and treatment purposes
  • A detection method for microbial drug resistance genes for non-disease diagnosis and treatment purposes

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Embodiment 1

[0029] The drug resistance gene detection of embodiment one human feces microorganism

[0030] Human fecal microbes reflect the types and quantities of human intestinal pathogenic microorganisms, and are the basis for the diagnosis and treatment of intestinal diseases. The human feces used in this example were taken from a patient with enteritis diagnosed by a doctor, and the detection of the drug resistance gene of microorganisms in the feces was to provide evidence for the doctor to use antibiotics to treat the patient's enteritis. This embodiment includes the following steps:

[0031] 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:

[0032] Among them, there is at least one drug resistance gene, at least one endogenous standard gene, and at least one exogenous stan...

Embodiment 2

[0063] Example 2: Detection of soil microbial drug resistance genes

[0064] 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.

[0065] The same drug resistance gene, endogenous standard gene and exogenous standard gene were determined by the same method as in Example 1, and the same multiple amplification primers and detection regions were obtained, which were produced by Tiangen Biochemical Technology (Beijing) Co., Ltd. The Soil Genome DNA Extraction Kit (Cat. No.: DP336) and the genome extraction of the mixed sample were carried out according to its operation manual. The total amount of the genome extracted from the mixed sample was 2350ng. After adding 2.350ng of exogenous nucleic acid in the same way as in Example 1, construct a h...

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Abstract

The invention discloses a method for detecting microbial drug resistance genes for non-disease diagnosis and treatment purposes. The method comprises: determining drug resistance genes, endogenous standard genes and exogenous standard genes; preparing multiple amplification primers; adding exogenous nucleic acid to the sample to be tested to obtain a mixed sample and extracting the genome; adding exogenous Standard genes, to obtain mixed nucleic acids; amplify the mixed nucleic acids, use the amplified products to construct a high-throughput sequencing library and perform high-throughput sequencing to obtain a sequenced fragment group; analyze the sequenced fragment group, and according to the obtained exogenous standard gene and The number of sequencing fragments of the endogenous standard gene is used to judge whether the experiment is successful; if the experiment is successful, the content of the drug resistance gene is calculated; according to the content of the drug resistance gene, it is determined whether the sample to be tested contains the drug resistance gene. The method can quantitatively detect any desired drug resistance genes in any microorganism at one time, and the detection does not need to cultivate and purify microorganisms, and the speed is fast, and the result is accurate and reliable.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting microbial drug resistance genes for non-disease diagnosis and treatment purposes. Background technique [0002] Antibiotics are widely used in the prevention and treatment of animal and plant diseases caused by microorganisms. Abuse of antibiotics will cause microorganisms to evolve resistance genes and produce drug resistance, which makes antibiotics ineffective in preventing and treating diseases. In order to effectively prevent To treat diseases caused by microorganisms and avoid the abuse of antibiotics, it is necessary to detect drug resistance genes for the samples to be tested. [0003] Existing methods for detecting drug-resistant genes include: estimating the pathogenic bacteria of the drug-resistant genes that need to be detected on the sample to be tested, isolating and purifying the pathogenic bacteria, and using PCR (Polymease Chain Reaction, polym...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2545/101C12Q2545/113C12Q2537/143
Inventor 彭海
Owner JIANGHAN UNIVERSITY