A kind of detection method of rice microbial drug resistance gene
A detection method, a technology for drug resistance
Active Publication Date: 2020-03-17
JIANGHAN UNIVERSITY
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Problems solved by technology
[0006] In order to solve the problem that the method for detecting drug-resistant genes in the prior art needs to be improved, the embodiment of the present invention provides a method for detecting drug-resistant genes of rice microorganisms
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Embodiment
[0030] Drug Resistance Gene Detection of Rice Microorganisms
[0031] The sample to be tested can be a whole rice plant or a part of the rice plant. In this embodiment, the sample to be tested is a rice leaf. The rice leaf used in this embodiment is taken from the rice field in Zhuankou Development Zone, Wuhan, and the resistance of microorganisms therein is detected. Drug-resistant genes are used to apply drugs to rice diseases in a targeted manner to avoid the emergence of drug resistance, and at the same time, reduce unnecessary drug use and save costs. 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 l...
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The invention discloses a method for detecting rice microbial resistance genes. 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 drug resistance genes of rice microorganisms. Background technique [0002] Antibiotics are widely used in the prevention and treatment of rice diseases. At present, in the process of rice disease prevention and control, the phenomenon of abuse of antibiotics is very common, which leads to the evolution of drug resistance genes and drug resistance of pathogenic microorganisms, making antibiotics effective in the prevention and treatment of rice diseases invalidated. Therefore, it is necessary to use targeted drugs against microbial resistance genes in rice to avoid drug resistance of pathogenic bacteria, and at the same time reduce the amount of pesticides and reduce costs. [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, iso...
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IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2545/101C12Q2545/113C12Q2537/143
Inventor 周俊飞陈利红彭海
Owner JIANGHAN UNIVERSITY