Test strip for identifying foot-and-mouth-disease virus infected and vaccine immunized animal at one step and preparation method of test strip
A technology for immunizing animals with foot-and-mouth disease virus, which is applied in the field of test strips for rapid identification of foot-and-mouth disease virus infection and vaccine immunization animals and the preparation thereof, can solve the problems of poor sensitivity, laboriousness, complicated operation and the like, and achieves high specificity, sensitivity, The results show the effect of intuitive, strong detection specificity
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[0051] 3. Preparation of Gold Standard SPA glass wool:
[0052] Preparation of gold sol by sodium citrate reduction method: add 2 to 4 ml of 0.5 to 2% trisodium citrate solution to 50 to 100 ml of boiling 0.01 to 0.05% chloroauric acid aqueous solution to obtain colloidal gold with a diameter of about 15 nanometers ; with K of 0.1mol / L 2 CO 3 Adjust the pH of the colloidal gold to 8.5-9.5, add the SPA to be labeled into the gold sol with a pH of 8.5-9.5 at a labeling ratio of 1:1000-1300, add 20% PEG10000 to the final concentration of 0.05% after labeling for 10 minutes , centrifuge at 4°C, 1500-3000 rpm for 20 minutes, remove unbound colloidal gold particles, centrifuge at 4°C, 15,000 rpm for 1 hour, discard the supernatant, and obtain colloidal gold-labeled SPA; 500 diluted colloidal gold-labeled SPA was adsorbed in refined glass wool, and dried under vacuum at 4°C to prepare gold-labeled SPA glass wool.
[0053] 4. The detection principle of the test strip of the present...
Embodiment 1
[0060] Embodiment one: one step distinguishes the test strip of foot-and-mouth disease virus infection and vaccine immune animal, see figure 1 , figure 2 In the figure, the support layer 1 is made of hard plastic thin strips, the sample adsorption fiber layer 2 is made of glass wool, the gold-labeled antibody fiber layer 3 is made of glass wool with colloidal gold-labeled SPA adsorbed, and the cellulose film layer 4 It is made of nitrocellulose membrane, and the water-absorbing material layer 5 is made of water-absorbing filter paper; the sample absorbing fiber layer 2, the gold-labeled antibody fiber layer 3, the cellulose film layer 4, and the water-absorbing material layer 5 are arranged from the left end (test end) to the right Paste on the hard plastic thin strips of the support layer 1 in turn, and the fibers at the junction between each other cross-penetrate each other; on the cellulose film layer 4, there are porcine foot-and-mouth disease virus structural protein VP...
Embodiment 2
[0062] Embodiment two: the test strip structure is basically the same as embodiment one, the difference is:
[0063] The gold-labeled antibody fiber layer 3 is made of glass wool adsorbed with colloidal gold-labeled SPA, and the detection blot 6-P printed with the porcine foot-and-mouth disease virus structural protein VP1 antigen solution expressed and purified by the Escherichia coli expression system is used for sheep (rabbit) The control blot C printed by the anti-SPA IgG solution, the combined blot band 7 formed by the arrangement of the two blots is "||” .
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