Simple method for detecting total content of human oral liquid immunoglobulin
An immunoglobulin and detection method technology, applied in the field of detection of total immunoglobulin content in human oral fluid, can solve the problems of long time, complicated operation, uneven viscosity of subjects, etc., and improve reliability and accuracy Effect
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Embodiment 1
[0073] A device for measuring the total immunoglobulin content of human oral fluid, comprising a human oral fluid collection tube, an oral fluid processing bottle, and colloidal gold detection test paper.
[0074] The human oral fluid collection tube is as figure 1 As shown, it includes the enrichment tube 1 and the drainage tube 2, the diameter of the drainage tube is smaller than the diameter of the enrichment tube, and the enrichment tube and the drainage tube are connected by a funnel-shaped connecting tube 3;
[0075] The diameter of the enrichment tube is 10-25mm, and the tube length is 30-75mm; the diameter of the drainage tube is 1-3mm, and the tube length is 20-50mm.
[0076] Preferably, the diameter of the enrichment tube is 12 mm, and the length of the tube is 40 mm; the diameter of the drainage tube is 2.5 mm, the length of the tube is 15 mm, and the length of the connecting tube is 10 mm.
[0077] The oral fluid treatment bottle is as figure 2 As shown, it incl...
Embodiment 2
[0085] The preparation scheme of antibody-coupled colloidal gold comprises the following steps:
[0086] Step (1): Add 1mL of 1% chloroauric acid solution to 100mL of deionized water, heat under stirring, and after boiling for 10-15 minutes, add 1% trisodium citrate solution to 0.01% chloroauric acid solution and stirring, the volume ratio of trisodium citrate solution to chloroauric acid solution is 1:40-1:50.
[0087] Step (2): Heat the solution obtained in step (1) to boiling. When the chloroauric acid solution turns dark red, keep boiling for 5 minutes, then stop heating, oscillate ultrasonically for 3-5 minutes, cool to room temperature, and place in Store in refrigerator at 4°C.
[0088] Step (3): the pH value of regulating colloidal gold solution is 7.5-8.5, just adds the monoclonal antibody A of mouse anti-human immunoglobulin (IgA, IgG and IgM three all can recognize) while stirring, after adding, Continue stirring for 20-30 minutes, then add 1× phosphate buffered s...
Embodiment 3
[0091] The preparation steps of the test paper for the semi-quantitative detection of the total immunoglobulin content in human oral fluid are as follows: the mouse anti-human immunoglobulin (IgA, IgG and IgM three can recognize) monoclonal antibody A coupled colloidal gold with 10 μ L / cm 2 Conjugate pad sprayed on glass cellulose membrane, marked; Goat anti-human immunoglobulin (IgA, IgG and IgM can recognize all three) polyclonal antibody B was sprayed on nitrocellulose membrane to form detection lines T1, T2 and T3 , the embedding amount is 0.01-1 μg and increases along the detection line from T1 to T3, wherein the embedding amount of T1 is 0.01-0.6 μg, the embedding amount of T2 is 0.01-0.3 μg more than that of T1, and the embedding amount of T3 The amount is 0.01-0.3 μg more than that of T2. The goat anti-mouse IgG antibody was sprayed on the nitrocellulose membrane to form a quality control line, and the embedding amount was 1-2 μg. Then the bottom plate, the sample pad...
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