Test strip for detecting citrobacter freundii and preparation method thereof
A kind of Freund's citric acid, anti-Freund's citric acid technology, applied in the field of immunochemical detection, can solve the problems such as the undisclosed Citrobacter freundii IgY, and achieve the effects of rapid detection, convenient storage, and low temperature requirements
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specific Embodiment 1
[0043] A test strip for Citrobacter freundii detection, such as figure 1 As shown, the test strip is composed of a PVC base plate 7 and a sample pad 1, a bonding pad 2, a nitrocellulose membrane 3, and a water-absorbing pad 4 that are sequentially overlapped on the PVC base plate 7; the bonding pad 2 is coated with anti-Freund’s lemon Acidobacillus egg yolk antibody-colloidal gold marker, nitrocellulose membrane 3 is respectively provided with detection line 5 coated with Citrobacter freundii ultrasonic disruption solution and quality control line coated with rabbit anti-chicken egg yolk antibody (IgY) 6.
[0044] The preparation method of the above-mentioned anti-Citrobacter freundii yolk antibody-colloidal gold marker is as follows: the anti-Citrobacter freundii yolk antibody and the colloidal gold solution with a particle diameter of 20-40nm are mixed at 5-7μg: 1mL, and mixed in Combine by stirring and shaking for 30-60min at pH 5.4, add PBST buffer containing 10wt% bovine...
specific Embodiment 2
[0045] The preparation method of quick detection Citrobacter freundii colloidal gold test strip, such as figure 2 As shown, the specific operation steps are as follows:
[0046] (1) Preparation of Citrobacter freundii
[0047] The preparation method of the Citrobacter freundii antigen is as follows: take the Citrobacter freundii strains stored in the laboratory at -80°C, streak and activate them in the agar medium, culture them upside down at 30°C for 24 hours, and pick a single colony to inoculate In a test tube containing 5mL beef extract-peptone liquid medium for seeds, cultivate at 30°C and 200rpm for 12-18h, then dilute the cultured bacteria liquid with beef extract-peptone liquid medium at 1:100 into the Erlenmeyer flask, and expand culture at 30°C and 200rpm , measure its OD600 value at regular intervals, stop culturing when its OD600 value reaches 0.4-0.6. Centrifuge the cultured bacterial solution at 5000rpm for 10min at 4°C, discard the supernatant, resuspend and ...
specific Embodiment 3
[0065] Colloidal gold test strips for the detection of clinical samples, the specific steps are as follows:
[0066] Various fish samples such as Chinese mitten crab, swimming crab, shrimp, grass carp, and crucian carp, as well as water samples from various local ponds were purchased from local supermarkets, and a total of 41 samples of their intestines, gills, and liver were dissected, and PBS buffer was added dropwise to On the tissue sample, break the tissue, centrifuge at 2000-3000rpm for 5 minutes, take the supernatant, insert the test strip into the sample to be tested, and observe the result after 10 minutes: if only a red line appears in the quality control area of the test strip It is a positive result, that is, there is Citrobacter freundii in the sample; if the detection line and the quality control line of the test strip have red lines at the same time, it is a negative result, that is, the sample does not contain Citrobacter freundii. Each sample was repeated 5 ...
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