Aeromonas hydrophila tetracycline drug-resistance gene detection method

Abstract

The invention discloses an aeromonas hydrophila tetracycline drug-resistance gene detection method. The method comprises the following steps: (1) collecting 0.5-0.7 g of aeromonas hydrophila living bacteria, resuspending with 5-7 mL of deionized water, repeatedly freeze-thawing to break cells, centrifuging and collecting a supernatant to obtain a template; (2) mixing the template with tetracycline drug-resistance gene detection primers TetA-F, TetA-R, TetC-F, TetC-R, TetG-F and TetG-R, and carrying out PCR amplification; and (3) carrying out agarose gel electrophoresis after PCR amplification in the step (2) and sending for sequencing. The detection method provided by the invention is sensitive, specific and rapid and can be adopted to detect a trace amount of target gene without the time-consuming aeromonas hydrophila culture stage.

Description

technical field [0001] The invention belongs to the technical field of pathogenic bacteria detection, and in particular relates to a method for detecting a drug-resistant gene of Aeromonas hydrophila tetracyclines. Background technique [0002] Tetracycline antibiotics are a class of broad-spectrum antibiotics produced by actinomycetes, including aureomycin, oxytetracycline and semi-synthetic derivatives methacycline, doxycycline, dimethylaminotetracycline, etc. This product is a broad-spectrum bacteriostatic agent, which has bactericidal effect at high concentrations. In addition to common Gram-positive bacteria, Gram-negative bacteria and anaerobic bacteria, most Rickettsia, Mycoplasma, Chlamydia, atypical mycobacteria, and spirochetes are also sensitive to this product. Due to the widespread use of tetracyclines over the years, most of the common clinical pathogens, including Gram-positive bacteria such as Staphylococcus and Gram-negative bacilli such as Enterobacter, ar...

AI Technical Summary

Problems solved by technology

The traditional drug susceptibility test must go through cumbersome steps such as separation and purification of Aeromonas hydrophila, reproduction and amplification, etc. The detection cycle is long, and the fastest it takes about 48 hours

Not conducive to timely drug selection and treatment

At the same time, the drug susceptibility test is to test the phenotypic drug resistance of Aeromonas hydrophila tentatively with drugs in vitro, and cannot detect the cryptic drug resistance of Aeromonas hydrophila

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