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A non-diagnostic detection method for microbial drug resistance genes in blood

A detection method, a drug resistance technology, applied in the biological field

Active Publication Date: 2020-01-31
JIANGHAN UNIVERSITY
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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 blood microbial drug resistance genes for non-diagnostic purposes

Method used

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  • A non-diagnostic detection method for microbial drug resistance genes in blood
  • A non-diagnostic detection method for microbial drug resistance genes in blood
  • A non-diagnostic detection method for microbial drug resistance genes in blood

Examples

Experimental program
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Embodiment

[0030] Drug resistance gene detection of human blood microorganisms

[0031] Human blood microorganisms reflect the types and quantities of pathogenic microorganisms for various diseases in the human body. The human cryopreserved blood samples used in this example were taken from cold patients. 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 method is 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 test sample. Exogenous standard genes are not present in organisms with known genomes.

[0034] The drug resistance gene in this example is resistant to penicillin...

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Abstract

The invention discloses a method for detecting blood microbial drug resistance genes for non-diagnostic 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 non-diagnostic method for detecting drug resistance genes of blood microorganisms. Background technique [0002] Antibiotics are widely used in the prevention and treatment of human diseases, and the abuse of antibiotics will cause microorganisms to evolve drug resistance genes and develop drug resistance, which makes antibiotics ineffective in preventing and treating diseases. Microbial drug resistance genes in blood are an important basis for drug use in many diseases. [0003] The existing methods for detecting drug resistance genes include: estimating the pathogenic bacteria of the drug resistance genes that need to be detected carried on the sample to be tested, isolating and purifying the pathogenic bacteria, using PCR (Polymease Chain Reaction, polymerase chain reaction) primers. The drug resistance gene is amplified, and electrophoresis or real-time quantitative PCR method...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2531/113C12Q2537/143C12Q2535/122C12Q2545/101
Inventor 胡长峰彭海
Owner JIANGHAN UNIVERSITY