Reagent case for diagnosing gene of pathogenic bacterial and para hematolysis vibrion of marine water product animal and hunman and testing method thereof
A technology for marine aquatic products and Vibrio hemolyticus, applied in biochemical equipment and methods, measurement/testing of microorganisms, and resistance to vector-borne diseases, etc., can solve problems such as limited application and development, troublesome preparation, and low sensitivity, and avoid Spread of germs, high practical value, and rapid effect
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Embodiment 1
[0031] Embodiment 1: the genetic diagnosis kit of Vibrio parahaemolyticus
[0032] The kit consists of the following parts (10 samples):
[0033] 1). Sample diluent (solution A), 1 tube, 5ml / tube, filled with 1×PBS, pH7.4.
[0034] 2).PCR. Reaction solution (solution B), 1 tube, 250μl / tube, containing PCR amplification reaction solution (25μl system), including ddH 2 O, containing Mg 2+ 10×Buffer, dNTP, primers VpF, VpR and TaqE.
[0035] 3).Positive control solution (solution C), 1 tube, 20μl / tube, containing the total DNA of Vibrio parahaemolyticus, as a positive template;
[0036] 4). Cuboid box, 8.5×5.8×6.2cm 3 .
[0037] 5). A piece of foam board, the same size as the bottom of the box, 2.2cm high, four rows of holes, four holes in the first row, hole diameter 1.3cm, five holes in the second row, hole diameter 1.0cm, third and fourth There are six holes in each row, and the hole diameter is 0.6cm. The above-mentioned small tubes are respectively placed correspondin...
Embodiment 2
[0048] Embodiment 2: Detection method of marine aquatic animals and human pathogenic bacteria-Vibrio parahaemolyticus
[0049] Use the test kit of embodiment 1, carry out according to the following steps:
[0050] 1). Using aseptic method, take 0.05 g of fresh abalone liver tissue, add 500 μl of sample diluent (liquid A) to dilute 10 times, and homogenize in an ice bath in a sterile homogenizer;
[0051] 2). Centrifuge at 6000r / min for 5min
[0052] 3). Take 100 μl of the supernatant, boil for 15 minutes, and immediately put it on ice for 5 minutes;
[0053] 4). Centrifuge at 6000r / min for 10min, and use the supernatant as a PCR template;
[0054] 5). Take 1 μl of the template and solution C respectively, add it to the PCR reaction solution (solution B), mix well, centrifuge at 1000r / min for 10sec, and place it on the PCR machine;
[0055] 6). Amplify according to the following conditions:
[0056] Pre-denaturation at 95°C for 3min→34 cycles at 94°C for 1min→10min at 72°C→st...
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