Vibrio shilonii multiple virulence factor GeXP rapid detection kit and detection method thereof
A technology of Vibrio schlossere and detection kit is applied in the field of rapid detection kits for pathogenic Vibrio-Vibrio schlosserii, can solve the problems of cumbersome process, time-consuming detection, low detection sensitivity, etc., and achieves wide detection coverage, The effect of improving accuracy
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Embodiment 1
[0056] (1) Genomic DNA extracted from Vibrio shilonii
[0057] Firstly, the Vibrio shilonii in the coral sample was purified and isolated on a plate, and then the Vibrio shilonii was cultured.
[0058] ① Add 50 μL of 10% (w / v) sterile Chelex-100 solution to the centrifuge tube;
[0059] ②Pick a single colony from the culture plate of Vibrio shilonii into the above Chelex-100 solution;
[0060] ③ Place the centrifuge tube on a vortex mixer and shake it fully for 10 seconds, take a boiling water bath for 10 minutes, cool to room temperature, and shake it fully for another 10 seconds;
[0061] ④ Centrifuge at 12000r / min for 2min, and the supernatant is the genomic DNA of Vibrio shilonii, and store it in a -20°C refrigerator for later use.
[0062] (2) Design and synthesis of primers for multigene PCR
[0063] Primers designed for the virulence gene sod (NCBI Reference Sequence: NZ_ABCH01000140.1), the amplified fragment size is about 110bp:
[0064] sod-F: AGGTGACACTATAGAATA ...
Embodiment 2
[0141] (1) Genomic DNA extracted from Vibrio shilonii
[0142] Firstly, the Vibrio shilonii in the coral sample was purified and isolated on a plate, and then the Vibrio shilonii was cultured.
[0143] ① Add 50 μL 10% (w / v) sterile Chelex-100 solution to the centrifuge tube;
[0144] ②Pick a single colony from the culture plate of Vibrio shilonii into the above Chelex-100 solution;
[0145] ③ Place the centrifuge tube on a vortex mixer and shake it fully for 10 seconds, take a boiling water bath for 10 minutes, cool to room temperature, and shake it fully for another 10 seconds;
[0146] ④ Centrifuge at 12000r / min for 2min, and the supernatant is the genomic DNA of Vibrio shilonii, and store it in a -20°C refrigerator for later use.
[0147] (2) Design and synthesis of primers for multigene PCR
[0148] Primers designed for the virulence gene sod (NCBI Reference Sequence: NZ_ABCH01000140.1), the amplified fragment size is about 110bp:
[0149] sod-F: AGGTGACACTATAGAATA CCG...
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