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Microorganism drug resistance gene detection method

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

Active Publication Date: 2016-05-11
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 microbial drug resistance genes

Method used

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  • Microorganism drug resistance gene detection method
  • Microorganism drug resistance gene detection method
  • Microorganism drug resistance gene detection method

Examples

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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 microorganism drug resistance gene detection method. The method includes the steps that a drug resistance gene, an endogenous standard gene and an exogenous standard gene are determined; multiple amplification primers are prepared; exogenous nucleic acid is added into a sample to be detected to obtain a mixed sample and extract a genome; the exogenous standard gene is added into the genome to obtain mixed nucleic acid; the mixed nucleic acid is amplified, a high-throughput sequencing library is constructed through amplification productimers, and high-throughput sequencing is conducted to obtain a sequencing fragment set; the sequencing fragment set is analyzed, and whether an experiment succeeds or not is judged according to the number of sequencing fragments of the obtained exogenous standard gene and the endogenous standard gene; if the experiment succeeds, the content of the drug resistance gene is calculated; according to the content of the drug resistance gene, whether the sample to be detected contains the drug resistance gene or not is judged. By means of the method, various drug resistance genes needing to be detected in any microorganism can be detected quantitatively at a time. Besides, microorganisms do not need to be cultured or purified during detection, the speed is high, and results are 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. 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 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 dr...

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 彭海
Owner JIANGHAN UNIVERSITY
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