Detection method of acidophilic thermophilic bacterium in fruit juice, and inducer and color development agent of detection method
A technology of acidophilic heat-resistant bacteria and detection method, which is applied in the field of detection of harmful microorganisms in fruit juice, can solve the problems of low detection rate, poor specificity and high cost, and achieve the effect of high sensitivity and specificity
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Embodiment 1
[0045] This example is a qualitative detection of thermostable acidophilus in fruit juice samples.
[0046] The fruit juice sample to be tested in this embodiment is 70°Brix concentrated apple juice that can be bought in a fruit juice company and is 8-12°Brix with sterile water.
[0047] The inducer is: 100 ml 402 medium + 0.17 g vanillic acid.
[0048] Induction culture: Add 1 ml of the fruit juice sample to be tested into the inducer at 45°C and culture it on a shaking table for 14 hours, then take 1 mL of the bacterial culture solution with a sterile pipette, centrifuge at 5000 rpm for 5 minutes, and take the supernatant.
[0049] The chromogenic reagent is: DNAzyme and 50 μL of 4 mM hydrogen peroxide. Among them, DNAzyme is 50 μL of 0.3 μM single-stranded nucleic acid sequence (5'-GTGGGTAGGGCGGGTTGG-3') (purchased from Sangon Bioengineering (Shanghai) Co., Ltd.), 50 μL of 1 μM hemin, and 100 μL of 20 mM potassium chloride solution, 100 μL of 150 mM sodium chloride soluti...
Embodiment 2
[0052] This example is the quantitative detection of thermostable acidophilus in fruit juice samples.
[0053] The fruit juice sample to be tested, the inducer, and the chromogen in this embodiment are the same as those in Example 1.
[0054] Make a standard curve between the concentration of acidophilus thermostable bacteria and the intensity of ultraviolet absorption:
[0055] (1) Prepare test strain samples
[0056] Aseptically operate, inoculate the strain of acidophilic thermostable bacteria (DSM3922, preserved in the German Culture Collection Center) into the Erlenmeyer flask, and culture it on a shaker at 45°C for 24 hours, as the test strain sample.
[0057] (2) Preparation of initial bacterial suspension
[0058] For aseptic operation, centrifuge the test strain sample obtained in step (1) at 5000 rpm for 5 min, discard the supernatant, and mix the bacteria with 10 mL of sterile water as a bacterial suspension.
[0059] Aseptic operation, pipette 1mL of bacterial s...
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