Four conjunction diagnostic kit of antimyocardial antibody
A diagnostic kit and myocardial antibody technology, which can be used in material analysis, measuring devices, instruments and other directions by observing the impact on chemical indicators, can solve the problems of lack of specificity and difficulty in clinical promotion of endomyocardial biopsy.
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Embodiment 1
[0093] Preparation of kit detection reagents
[0094] 1. Synthesis of Antigenic Peptides
[0095] Antigenic peptides with the same antigenicity as the four myocardial antigens were synthesized.
[0096] 1. The four myocardial antigen peptides and their amino acid sequences are:
[0097] ④ Cardiomyocyte mitochondrial inner membrane ADP / ATP carrier protein antigenic peptide (ANT);
[0098] ⑤β 1 Adrenergic receptor antigen peptide (β 1 );
[0099] ⑥ M 2 - Cholinergic receptor antigenic peptides (M 2 );
[0100] ④ Myosin heavy chain antigen peptide (MHC).
[0101] Amino acid sequence of ADP / ATP carrier protein peptide:
[0102] ANT-1 Pro-Ile-Glu-Arg-Val-Lys-Leu-Leu-Leu-Gln
[0103] ANT-2 Tyr-Asp-Glu-Ile-Lys-Lys-Phe-Val
[0104] beta 1 Amino acid sequence of adrenergic receptor peptide:
[0105] His-Trp-Trp-Arg-Ala-Glu-Ser-Asp-Glu-Ala-Arg-Arg-Cys-Tyr-Asn-Asp-Pro-Lys-Cys-Cys-Asp-Phe-Val-The-Asn- Arg-Cys
[0106] m 2 -Amino acid sequence of cholinergic receptor peptid...
Embodiment 2
[0141] Operation procedure using the diagnostic kit of the present invention
[0142] 1. Prepare washing liquid: take an appropriate amount of concentrated washing liquid and dilute it with distilled water according to the ratio of 1:9 (volume ratio) for later use;
[0143] 2. Add sample diluent: add 100 μl to each reaction well with a sampler;
[0144] 3. Add samples and leave blank control: Add 1 μl of each sample to be tested into each well of the existing sample diluent, and leave one well without adding a sample as a blank control.
[0145] 4. Add negative control: Add negative control serum to the 2 wells set as negative control, add 1 μl to each well;
[0146] 5. Mix well and incubate in a 37°C incubator for 60 minutes;
[0147] 6. Wash the plate: Pour off the contents, fill the reaction well with washing solution, let it stand for 3 minutes, pat dry and repeat this 3 times;
[0148] 7. Add enzyme-labeled secondary antibody: add 100 μl enzyme-labeled secondary antibo...
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