Quantitative detection method of resistance genes of intestinal bacterial antibiotics

A quantitative detection method and technology of antibiotic resistance, applied in the field of quantitative detection of antibiotic resistance genes of intestinal bacteria, can solve the problems of underestimating the number of intestinal microorganisms and the deviation of the carrying amount of intestinal bacteria resistance genes, and achieve high theoretical research. Significance and practical value, stable and reliable results, accurate test results
CN107254518AInactive Publication Date: 2017-10-17SUN YAT SEN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
SUN YAT SEN UNIV
Publication Date
2017-10-17
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a quantitative detection method of resistance genes of intestinal bacterial antibiotics; the method comprises the steps of S1, extracting total DNA of intestinal bacteria; S2, using the total DNA of the intestinal bacteria as a template, amplifying antibiotic resistance genes and 16SrRNA gene via PCR (polymerase chain reaction), and linking the amplification products to cloning vectors, and converting host bacteria to obtain positive clones; propagating the positive clones, and extracting plasmids, measuring copy density of the plasmids, diluting at 100 gradient to obtain a standard template, and establishing a standard curve for the copy number and Ct value through fluorescent quantitative PCR; S3, subjecting the positive plasmids and samples to fluorescent quantitative PCR amplification, and calculating the copy number of antibiotic resistance genes carried in the samples to obtain relative content of the antibiotic resistance genes. The method is good in safety and environmental friendliness and low in price, has simple steps, is low in time consumption, has stable and reliable results, has good repeatability, high sensitivity and high practicality, is simple and fast, and has a promising application prospect.
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Description

technical field

[0001] The invention belongs to the field of biotechnology. More specifically, it relates to a quantitative detection method for antibiotic resistance genes of intestinal bacteria. Background technique

[0002] In recent years, due to the extensive use of antibiotics in aquaculture and animal husbandry, the problem of environmental pollution has become increasingly serious, and it has become one of the current international research hotspots. my country is a big country for Litopenaeus vannamei farming, but in recent years, due to the aggravation of aquaculture pollution, the quality of the aquaculture environment has continued to deteriorate, the variety of pathogenic organisms has increased and the speed of transmission has accelerated, and aquaculture diseases have become increasingly serious. The disease problems in aquaculture are especially bacterial diseases. It occurs frequently and seriously in intensive farming systems, which greatly limits the deve...

Claims

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