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Method for detecting drug-resistant genes of pathogenic microorganisms of human body

A technology of pathogenic microorganisms and detection methods, applied in the field of detection of drug resistance genes of human pathogenic microorganisms

Active Publication Date: 2016-06-08
JIANGHAN UNIVERSITY +1
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  • Summary
  • 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 human pathogenic microorganisms

Method used

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  • Method for detecting drug-resistant genes of pathogenic microorganisms of human body
  • Method for detecting drug-resistant genes of pathogenic microorganisms of human body
  • Method for detecting drug-resistant genes of pathogenic microorganisms of human body

Examples

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Embodiment 1

[0029] Example 1. Drug resistance gene detection of human feces microorganisms

[0030] The sample to be tested can be human organs and tissues, a part of human organs and tissues, human body fluid, human secretion or human excrement, and in this embodiment, the sample to be tested is human feces (human excrement), and human feces microbial reaction The types and quantities of pathogenic microorganisms in the human intestinal tract are determined, which is 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...

Embodiment 2

[0063] Embodiment two, the drug resistance gene detection of human blood microorganism

[0064] Human blood microbes reflect the types and quantities of pathogenic microorganisms in various diseases of the human body, and are the basis for disease diagnosis and treatment. The cryopreserved human blood samples used in this example are taken from cold patients, and the detection of the drug resistance genes of microorganisms in the blood is to provide evidence for doctors to use antibiotics for treatment. The steps of this implementation are roughly the same as those in Embodiment 1, and the following focuses on the differences:

[0065] By the same method as in Example 1, determine the drug resistance gene of the microorganisms 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 multiple amplification primers of the test regions of sex genes, endogenous standard gene...

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Abstract

The invention discloses a method for detecting drug-resistant genes of pathogenic microorganisms of a human body. The method comprises the steps that drug-resistant genes, endogenous standard genes and exogenous standard genes are determined; multiple amplification primers are prepared; exogenous nucleic acid is added to a sample to be detected, a mixed sample is obtained, and a genome is extracted; the exogenous standard genes are added to the genome, and mixed nucleic acid is obtained; the mixed nucleic acid is amplified, the amplification product is utilized for constructing a high-throughput sequencing library, high-throughput sequencing is carried out, and a sequencing fragment set is obtained; the sequencing fragment set is analyzed, and whether an experiment succeeds or not is judged according to the number of the obtained sequencing fragments of the exogenous standard genes and the endogenous standard genes; if the experiment succeeds, the content of the drug-resistant genes is calculated, and whether the sample to be detected contains the drug-resistant genes or not is judged according to the content of the drug-resistant genes. According to the method, any multiple drug-resistant genes expected to be detected in any microorganism can be quantitatively detected at a time, the microorganisms do not need to be cultured or purified during detection, the speed is high, and the result is accurate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting drug resistance genes of human pathogenic microorganisms. Background technique [0002] Antibiotics are widely used in the prevention and treatment of human diseases. Abuse of antibiotics will lead to the evolution of resistance genes and drug resistance in microorganisms, which makes antibiotics ineffective in the prevention and treatment of diseases. Drug resistance genes of pathogenic microorganisms are an important basis for drug use in the treatment of various diseases. [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 P...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2537/143C12Q2535/122C12Q2545/101C12Q2545/113
Inventor 张静张英彭海卢龙
Owner JIANGHAN UNIVERSITY
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