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Method for detecting resistance gene tetG in offshore sea area

A technology of resistance gene and detection method, which is applied to the detection field of resistance gene tetG in offshore waters, can solve the problem of inability to accurately quantify the resistance gene tetG and the like, and achieve the effect of improving the accuracy

Inactive Publication Date: 2021-02-02
TIANJIN UNIV
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Problems solved by technology

[0005] 2. Solve the problem that the non-culturable resistance gene tetG in seawater cannot be accurately quantified when detecting resistance genes based on selective culture methods

Method used

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  • Method for detecting resistance gene tetG in offshore sea area
  • Method for detecting resistance gene tetG in offshore sea area
  • Method for detecting resistance gene tetG in offshore sea area

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Embodiment Construction

[0027] The method established in this study is suitable for the quantification of the resistance gene tetG in seawater samples. The method specification is as follows:

[0028] 1. At the set sampling point, collect water samples about 0.5m below the water surface with a special sample collector for water samples, put them into sterile glass bottles with a volume of 1L, and store them in ice boxes after numbering. After the sample collection is completed, it is immediately transported back to the laboratory at a low temperature of 4°C for processing. After transporting back to the laboratory, some water samples are filtered with a microporous filter membrane with a pore size of 0.22 μm. Each filter membrane filters 500 mL of water samples. Filter three filters per sampling point and store at -80°C until subsequent DNA extraction.

[0029] 2. Extract sample DNA. E.N.Z.A.TM Water DNA Kit (Omega, USA) is used for DNA extraction of water samples. In order to ensure the elimination ...

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Abstract

The invention discloses a method for detecting a resistance gene tetG in an offshore sea area. The method comprises the following steps of extracting total DNA of an environmental sample; amplifying atarget fragment; constructing a standard plasmid template; establishing a Q-PCR quantitative standard curve; and determining the resistance gene tetG of the environmental sample. Compared with a detection method based on selective culture at present, the method has the following advantages that firstly, the detection time is shortened from 3-4 days of the original method to 1 day, and only 2-3 hours are needed under the condition that the standard curve is completed; and secondly, the problem that the resistance gene tetG carried by uncultured microorganisms cannot be determined by the original method is solved, and the identification accuracy is improved.

Description

technical field [0001] The invention relates to the technical field of common gene detection, in particular to a method for detecting resistance gene tetG in offshore waters. Background technique [0002] Toxic chemicals flowing into the ocean during development in coastal areas pose health risks to humans and ecosystems, and antibiotic resistance genes (ARGs) have also been identified as important pollutants. Antibiotic resistance genes (ARGs) are an emerging class of pollutants that have attracted great attention due to their potential ecotoxicological risks and threats to public health. Among them, tetracycline resistance genes have attracted much attention due to their wide range of uses, high excretion rate, high solubility and high persistence in the environment. exist in both, so a simple and rapid method is needed for rapid and accurate detection of the tetracycline resistance gene tetG. Contents of the invention [0003] The purpose of the invention is to solve ...

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Application Information

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IPC IPC(8): C12Q1/6851
CPCC12Q1/6851
Inventor 汪光义郭嘉懿李佳倩何艺科
Owner TIANJIN UNIV
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