Sensitization blocking buffer solution and application thereof
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A buffer and buffer salt technology, applied in the field of sensitization blocking buffer, can solve the problems of unsatisfactory sensitivity, cross-interference, test specificity, etc., to improve sensitivity and specificity, improve P/N , the effect of solving false positives
Active Publication Date: 2020-05-22
深圳爱信生物技术有限公司
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Problems solved by technology
At present, no Helicobacter pylori antibody detection kit has been used clinically. One of the main reasons is that the sensitivity of conventional detection methods cannot meet the requirements due to the low antibody content in urine, and the presence of other interfering substances is easy to form cross-interference, which is harmful to the test. influence of specificity
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Embodiment 1
[0023] Example 1. Preparation and detection of Helicobacter pylori urine antibody detection kit (colloidal gold method) 1. Preparation of detection kit and detection of urine specimen
[0024] 1. Preparation of colloidal gold particles: Take 800mL of purified water in a round bottom flask and place it in a magnetic stirring heating mantle, then add 8mL of 1% (w / v) HAuCl 4 , stir and heat, after the solution boils, quickly add 15mL trisodium citrate with a concentration of 1% (w / v), continue stirring and heating for 10 minutes to obtain a wine red colloidal gold solution, stop heating and continue stirring to cool to room temperature, filter and store Store in a clean glass bottle;
[0025] 2. Colloidal gold labeled goat anti-human IgG: measure 100mL colloidal gold in a clean beaker, stir well, use 0.2MK 2 CO 3 Adjust the pH to 9.0, then slowly add 0.5 mg of goat anti-mouse IgG dropwise, and react for 10 minutes; then slowly add 10 mL of 10% (w / v) BSA, stir and react for 10 m...
Embodiment 2
[0043] Example 2, Preparation of Helicobacter pylori urine antibody detection kit (enzyme-linked immunoassay) and detection of urine specimen
[0044] 1. Coating of 96-well plate (Guangzhou Jiete Company, high binding force)
[0045] Take 100 μg Helicobacter pylori cytotoxin-associated protein A (CagA) recombinant protein, 50 μg Helicobacter pylori vacuolating toxin (VacA) recombinant protein, and 50 μg Helicobacter pylori urease (Urease) recombinant protein and add it to 100 mL carbonate buffer (0.05 M CB, pH 9.6), mixed thoroughly, coated in a 96-well plate, each well was coated with 100 μL, put the coated plate in the refrigerator, and coated overnight at 2-8°C.
[0046] 2. The coating is closed by the board
[0047] Wash the plate twice with washing solution (0.01M PBS, containing 0.05% Tween20, pH7.2), add 150 μL of blocking solution to each well to block, and block at room temperature (20-28°C) for 1 hour. The components of blocking solution are: 1% (w / v) BSA, containi...
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Abstract
The invention belongs to the technical field of diagnostic reagents, and discloses a sensitization blocking buffer solution containing a buffer solution, 0.2%-0.2% 10% (v/v) ethanol; 0.1%-5%(w/v)PEG6000, 0.01%-0.5% (w/v) of sodium carboxymethyl cellulose (M.W.90000), 0.005%-0.25% (w/v) of poloxamer 188, 0.05%-5% (w/v) of glycine betaine, 0.1% (v/v) of Tween20 and 0.045% (v/v) of ProClin 300. The buffer solution is a phosphate buffer salt solution with the pH value of 5.8-8.0, an HEPES buffer salt solution with the pH value of 6.8-8.2, a PIPES buffer salt solution with the pH value of 6.1-7.5,an MOPS buffer salt solution with the pH value of 6.5-7.9 and a Tris buffer salt solution with the pH value of 7.1-9.0. The sensitization blocking buffer solution and a to-be-detected sample are incubated at the same time to participate in reaction; the background of a negative sample of a detection result is remarkably reduced, the signal value of a positive sample detection result is increased,the ratio of the positive sample signal value to the negative sample signal value of the detection result is increased under the dual action of blocking non-specific and sensibilization specific reactions, and the detection sensitivity and specificity are improved. The sensitization blocking buffer solution can be used for preparing a helicobacter pylori urine antibody detection kit.
Description
technical field [0001] The invention belongs to the technical field of diagnostic reagents, and in particular relates to a sensitization blocking buffer, a kit and an application. Background technique [0002] Immunodiagnosis (immunodiagnosis) is the application of the theory, technology and methods of immunology to diagnose various diseases and determine the immune status. Immunodiagnostic reagents are widely used in hospitals, third-party testing, blood stations, and physical examination centers. They are mainly used for infectious disease detection, infectious disease detection, tumor detection, and metabolic marker detection. Among them, immunodiagnosis includes radioimmunoassay, enzyme-linked immunoassay, immunochromatography, chemiluminescence, etc. Immunodiagnosis has the following characteristics: ①Strong specificity, try to avoid cross-reaction and false positive, so as to ensure the accuracy of diagnosis; ②High sensitivity, can detect trace reaction substances and...
Claims
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Application Information
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