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Amoxicillin antibody detection kit and preparation and application method thereof

An amoxicillin and antibody detection technology, applied in biological testing, measuring devices, material testing products, etc., can solve the problems of insufficient guarantee of the efficacy of amoxicillin drugs, endangering the life safety of patients, affecting the treatment effect, etc. The effect of strong sensitivity and stability, convenient clinical application and simple operation

Inactive Publication Date: 2018-11-20
江苏中济万泰生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Once the patient develops anti-amoxicillin antibodies, the antigen-antibody complex formed by the antibodies and amoxicillin drugs will neutralize and inhibit the efficacy of amoxicillin drugs, so that the efficacy of amoxicillin drugs cannot be fully guaranteed
At the same time, the combination of the immune complex formed by the amoxicillin drug and its corresponding antibody and human red blood cells will cause red blood cell hemolysis, which will affect the treatment effect in the slightest, delay the disease, and endanger the life of the patient in severe cases.

Method used

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  • Amoxicillin antibody detection kit and preparation and application method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] This embodiment relates to the preparation of amoxicillin antibody detection kit (column agglutination method)

[0060] Step 1. Amoxicillin Antibody Detection Reagent Card

[0061] (1.1) Measuring: Receive the dextran gel (swelling gel) in batches, measure and transfer the dextran gel (swelling gel) to a clean and sterilized container.

[0062] (1.2), washing: wash the dextran gel (swelling gel) with 0.9% sodium chloride solution, the volume ratio of 0.9% sodium chloride solution to dextran gel (swelling gel) is 2:1, mix well After centrifuging at 2000rpm for 1 minute, the supernatant was removed, and the operation was repeated 5 times to obtain a washed gel for marking.

[0063] (1.3), preparation: measure antihuman globulin reagent (anti-IgG+C 3 d). Anti-human globulin reagent (anti-IgG+C 3 d) Add to the washed gel and mix, and mix evenly with utensils to form an antibody gel, and mark it for use.

[0064] (1.4), filling, centrifugation: adjust the continuous sam...

Embodiment 2

[0082] This embodiment relates to the preparation method of amoxicillin monoclonal antibody

[0083] (1) Preparation of feeder cells:

[0084] Take Balb / c mice aged 6-10 weeks, kill them by pulling their necks, soak them in 75% alcohol, and disinfect them for 3-5 minutes; transfer them to an aseptic operating table, and cut the skin with sterile scissors to fully expose the peritoneum; Inject 10 mL of pre-cooled serum-free 1640 culture solution into a sterile syringe, gently tap the abdominal cavity of the mouse with the bottom of the tweezers, suck out the culture solution, put it into a 10 mL centrifuge tube, and centrifuge at 1200 r / min for 10 min; discard the supernatant, and use complete culture solution Resuspend the feeder cells, count, and adjust the number of cells to 2×105 / mL, add the feeder cells to a 96-well plate, 100 μL per well, and then culture in a 37°C CO2 incubator;

[0085] (2) Hybridoma cell fusion and subcloning screening by limiting dilution method

[...

Embodiment 3

[0100] This embodiment relates to the use of amoxicillin antibody detection kit

[0101] 1. Take the amoxicillin antibody detection reagent card, and mark 4 wells (I, II, III, and IV) for each patient sample to be tested.

[0102] 2. Add 25 μl of amoxicillin-treated erythrocyte suspension to each well of wells I, II, and III, and add 25 μl of non-amoxicillin-treated erythrocyte suspension to well IV.

[0103] 3. Add 50 μl of plasma of the sample to be tested to wells I and IV respectively, add 50 μl of positive control solution to well II, and add 50 μl of negative control solution to well III. Mix well and incubate for 1 hour at 37±1°C in a reagent card incubator.

[0104] 4. Centrifuge in LB-3000 medical centrifuge for 5min (900rpm×2min, 1500rpm×3min), observe and record the results within 30min.

[0105] 5. Result judgment

[0106] Positive result: Red blood cells located on the surface of the gel or suspended in the gel are positive reactions.

[0107] Negative result:...

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Abstract

The invention relates to an amoxicillin antibody detection kit and a preparation and application method thereof. The amoxicillin antibody detection kit is mainly composed of an amoxicillin antibody detection reagent card, amoxicillin processing erythrocyte, non-amoxicillin processing erythrocyte, an amoxicillin antibody positive contrast liquid, and an amoxicillin antibody negative contrast liquid. The kit has simple in operation, is strong in sensitivity and stability, and is more convenient in a clinical application, and can rapidly detect an amoxicillin antibody for a patient.

Description

technical field [0001] The invention belongs to the field of antibiotic antibody detection, and in particular relates to a detection kit, a preparation method and a use method of amoxicillin antibody. Background technique [0002] With the advancement of science and technology and the development of the pharmaceutical industry, more and more antibiotics are used clinically, resulting in the abuse of antibiotics, making the clinical use of antibiotics ineffective and even hemolysis in patients. In the slightest, it will affect the treatment effect and delay the treatment, and in severe cases, it will endanger the life safety of patients, cause medical accidents, and cause disputes between doctors and patients. Up to now, there is no effective method to detect antibiotic antibodies. In order to avoid the hemolytic side effects of antibiotics, multiple antibiotics can only be used simultaneously for treatment. Therefore, there is an urgent clinical need for antibiotic antibody...

Claims

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Application Information

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IPC IPC(8): G01N33/50
CPCG01N33/5052
Inventor 赵树铭林裕翔吴明磊
Owner 江苏中济万泰生物医药有限公司
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