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Kit for detecting autoimmune liver disease related antibody autoantibody to nuclear antigen (ANA) and detection method thereof
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An autoimmune, kit-based technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as differences between new and old tissue slices, and difficulty in developing
Inactive Publication Date: 2013-03-06
AUTOBIO DIAGNOSTICS CO LTD
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Clinically, indirect immunofluorescence (IIF) is commonly used to detect ANA in patients. The experimental matrix is usually frozen sections of liver, stomach, and kidney tissues of different primates, which contain a complete antigen spectrum. The identification of staining has a certain degree of subjectivity, so the interpretation of the results may vary at different time points, between different observers, and between old and new tissue sections, and requires the assistance of fluorescent microscopes and other instruments. In general small and medium hospitals Difficult to carry out
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Embodiment 1
[0021] The kit (plate type) for detecting autoimmune liver disease-related antibody ANA described in the present invention includes:
[0022] 1. Preparation of target antigen-carrier conjugate: Add 100 μl of target antigen (natural antigen, recombinant antigen, synthetic peptide, HepG2 cell, Hep2 cell, Helacell or cell lysed antigen) not higher than 10ug / ml to a 96-well plate In the medium, coat at 4°C for 16-24h or at 37°C for 2h, then wash twice with the prepared (PBST) washing solution;
[0023] 2. Blocking: Use 50-200 μl of 0.01-0.1M phosphate buffer containing 0.05-0.5% Tween20, 1-5% BSA, 1-5% Casein, 1-5% peptone, 5-10% sucrose, Block at 4°C for 16-24h or at 37°C for 2h, then shake off excess buffer, place in a dry environment to dry, and seal at 2-8°C for storage;
[0024] 3. Preparation of calibrator, positive control substance and negative control substance: take a clinically determined high-value sample, dilute it with the calibrator diluent (PBS+3%BSA) at a rate o...
Embodiment 2
[0039] The kit (magnetic particles) for detecting autoimmune liver disease-associated antibody ANA described in the present invention includes:
[0040] 1. Preparation of target antigen-carrier conjugates: Take 10mg of magnetic particles, wash them with phosphate buffer for 5 times, add no less than 0.15ml of EDC or (and) NHS or glutaraldehyde (and) activator for 2 hours, wash 3 times, then add 0.1mg of target antigen, shake at room temperature for 2h;
[0041] 2. Blocking: Use 1ml of 0.01-0.1M phosphate buffer containing 0.05-0.5% Tween20, 1-5% BSA, 1-5% Casein, 1-5% peptone, 5-10% sucrose, shake at room temperature Block for 10 minutes, block four times in a row, then add 2ml of blocking solution and store at 2-8°C;
[0042] 3. Preparation of calibrator, positive control substance and negative control substance: take a clinically determined high-value sample, dilute it with the calibrator diluent (PBS+3%BSA) at a rate of not less than 1:50, and take a linear comparison The...
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Abstract
The invention discloses a kit for detecting an autoimmune liver disease related antibodyautoantibody to nuclear antigen (ANA). The invention also discloses a method for quantitatively and qualitatively detecting ANA by using the kit. The method comprises the following steps of: combining a detection sample with a specific antigen which corresponds to a marker-linked anti-human IgG or / and anti-human IgM or / and anti-human IgA and a detection antibody, adding substrates of a luminous system or a developing system or a light exciting system, reading a signal value of a sample to be detected, calculating a quantitative and qualitative result, and reading the sample through immunochromatography or immuno-infiltration naked eyes or instruments to obtain a quantitative and qualitative result, thus accurately diagnosing the type of severity of the autoimmune liver disease, and guiding the medication of a doctor. The kit and the detection method have high sensitivity, high specificity and high accuracy and are suitable for semi-automatic or full-automatic detection systems or naked eye interpretation.
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