Quantitative detection method for sulfonamide-antibiotic-resistant genes in seawater bathing spot

A quantitative detection method and antibiotic resistance technology, applied in the field of detection, can solve the problems of late resistance genes, little attention, serious problems, etc., and achieve the effect of high amplification efficiency, high measurement accuracy and wide application range

Pending Publication Date: 2020-06-19
NATIONAL MARINE ENVIRONMENTAL MONITORING CENTRE
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Problems solved by technology

Therefore, the pollution of sulfa resistance genes caused by sulfa antibiotics is becoming more and more serious
However, the research on the distribution of resistance genes in bathing beaches started late in China, and less attention has been paid to them. At present, a unified detection method has not been established to detect antibiotic resistance genes in bathing beaches.

Method used

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  • Quantitative detection method for sulfonamide-antibiotic-resistant genes in seawater bathing spot
  • Quantitative detection method for sulfonamide-antibiotic-resistant genes in seawater bathing spot
  • Quantitative detection method for sulfonamide-antibiotic-resistant genes in seawater bathing spot

Examples

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Embodiment Construction

[0040] This example specifically illustrates the quantitative detection method for determining the resistance genes sul1 and sul2 of sulfonamide antibiotics in a bathing beach.

[0041] Step 1: Collection method of water samples and sediments in beach

[0042] Water samples: Water samples were collected in sterilized 5L polyethylene buckets. It is advisable to take surface water when collecting, and collect 3 barrels at each sampling point as 3 samples in parallel, and mark the serial number, collection location, and collection time. Immediately after collection, put the water sample into an incubator filled with ice cubes, and bring it back to the laboratory for processing in time.

[0043] Sediment: The surface sediment is collected with a sterilized sampler and put into a sterile ziplock bag. Immediately after collection, the samples were placed in an incubator filled with ice cubes and brought back to the laboratory for processing in time.

[0044] Step 2: Sample transp...

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Abstract

The invention discloses a quantitative detection method for sulfonamide-antibiotic-resistant genes in a seawater bathing spot. The method comprises the steps of subjecting a water sample and depositsof the seawater bathing spot to collecting and low-temperature storage, carrying out DNA extraction and purification on the collected sample, then, amplifying DNA fragments of the antibiotic-resistantgenes, cloning the DNA fragments to a host cell to construct standard plasmids, preparing a fluorescent quantitative PCR standard curve of a target DNA standard substance, and calculating the contentof the antibiotic-resistant genes in the sample according to the PCR standard curve of the standard substance and a CT value of the purified sample. According to the method disclosed by the invention, the steps are simple, the operation is convenient, a sample DNA sequence analyzing step is omitted, and the content of the antibiotic-resistant genes in the sample water and the deposits is obtainedaccording to the PCR standard curve and the CT value of the sample, so that the method is wide in applicable range.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to a quantitative detection method for sulfonamide antibiotic resistance genes in bathing beaches. Background technique [0002] With the increasing development of coastal tourism resources and the continuous increase of coastal population, bathing beaches have gradually become one of the important media for the spread of antibiotic resistance genes. Antibiotic resistance genes will be brought into nearshore seaside tourist areas along with rainwater erosion, tides and ocean currents. Through the transfer of genetic material between bacteria species, antibiotic resistance genes may be introduced into human pathogenic bacteria in seawater environment. Because the beach entertainment waters carry a large number of tourists, these antibiotic resistance genes can cause fatal intestinal diseases among swimmers swallowed by seawater, thus greatly increasing the possibility o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6895C12Q1/686
CPCC12Q1/689C12Q1/6895C12Q1/686C12Q2563/107C12Q2545/114C12Q2561/113
Inventor 苏洁樊景凤明红霞付韵涵赵小慧石婷婷金媛
Owner NATIONAL MARINE ENVIRONMENTAL MONITORING CENTRE
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