Bivalent gold-labeled test strip for classical swine fevervirus and porcine pseudorabies virus and preparation method thereof
A technology of porcine pseudorabies virus and swine fever virus, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of systemic neurological symptoms, high detection cost, and long time consumption
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Embodiment 1
[0074] Such as Figure 1-2 As shown, a double-linked gold standard test paper for swine fever virus and porcine pseudorabies virus includes a support plate 1 and an adsorption layer fixed on the support plate. The adsorption layer is sample pad 3 and gold standard pad in sequence from the test end , NC film 2 and water-absorbing pad 6, the gold standard pad comprises the upper strata gold standard pad 4 (being gold standard pad 2) and the lower floor gold standard pad 5 (being gold standard pad 1) stacked up and down arranged on the supporting plate successively, Colloidal gold-labeled anti-swine fever virus monoclonal antibody mAb1 is adsorbed on the pad, colloidal gold-labeled anti-pseudorabies virus monoclonal antibody mAb2 is adsorbed on the upper gold pad, and rabbit anti-mouse IgG antibody or The quality control line (C) 7 printed by Staphylococcus aureus SPA, the detection line (T1) 8 printed by anti-CSFV monoclonal antibody mAb3, and the detection of anti-porcine pseud...
Embodiment 2
[0079] The preparation of the double-linked gold standard detection test paper of swine fever virus and porcine pseudorabies virus of the present invention: firstly prepare the immunogen of swine fever virus and porcine pseudorabies virus, and then prepare and screen the corresponding high-affinity and high-specificity monoclonal antibodies . Among them, monoclonal antibody mAb1 of classical swine fever virus and mAb2 of monoclonal antibody of porcine pseudorabies virus are used to prepare gold-labeled antibodies, and mAb3, a monoclonal antibody that specifically recognizes classical swine fever virus and is different from the epitope recognized by mAb1, is used to print classical swine fever virus Detection line blot T1 "│", the monoclonal antibody mAb4 that specifically recognizes porcine pseudorabies virus and has a different epitope from mAb2 is used to print porcine pseudorabies virus detection line blot T2 "│"; secondly, prepare rabbit anti-mouse IgG antibody or Staphylo...
Embodiment 3
[0141] Embodiment 3, the specificity of test paper, sensitivity detection
[0142] Specificity: For test paper specificity test results, see Figure 7 , the test paper has no cross-reaction with PRRSV, PEDV, PCV2 and other porcine viruses.
[0143] Sensitivity: The sensitivity test of the test paper uses CSFV and PRV virus cultured by cells for detection, and the sample uses the PBS dilution method to dilute the CSFV or PRV virus titer to 10 respectively. 6 TCID50 / mL, in addition, add PRV virus or CSFV virus to the same buffer for 2-fold dilution for sensitivity testing. The results of the visual sensitivity of the dual test paper show that the test paper has a concentration of CSFV ≥ 10 4.6 TCID50 / mL and PRV is 10 6 When TCID50 / mL, the test paper shows a positive result, and when the PRV concentration is ≥ 10 3.2 TCID50 / mL and CSFV are 10 6 The test paper showed a positive result when TCID50 / mL, so the sensitivity of the test paper to CSFV and PRV were 10 4.6 TCID50 / mL ...
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