Ampicillin antibody detection kit and preparation and use methods thereof

A technology for ampicillin and antibody detection, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of insufficient guarantee of the curative effect of ampicillin, endanger the life safety of patients, affect the treatment effect, etc., and achieve sensitivity and stability Strong, convenient clinical application, simple operation

Inactive Publication Date: 2018-09-28
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 a patient develops anti-ampicillin antibodies, the antigen-antibody complex formed by the antibody and ampicillin will neutralize and inhibit the efficacy of ampicillin, so that the efficacy of ampicillin cannot be fully guaranteed
At the same time, the combination of the immune complex formed by ampicillin and its corresponding antibody and human red blood cells will cause hemolysis of red blood cells, which will affect the treatment effect in the slightest, delay the disease, and endanger the life safety of the patient in severe cases.

Method used

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  • Ampicillin antibody detection kit and preparation and use methods thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0060] Step 1. Ampicillin 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 sampl...

Embodiment 2

[0082] This embodiment relates to the preparation method of ampicillin 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 put them in 37°C CO 2 incubator cultivation;

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

Embodiment 3

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

[0101] 1. Take the ampicillin 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 ampicillin-treated erythrocyte suspension to each of wells I, II, and III, and add 25 μl of non-ampicillin-treated erythrocyte suspension to well IV.

[0103] 3. Add 50 μl of the sample plasma 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: all red blo...

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Abstract

The invention relates to an ampicillin antibody detection kit and preparation and use methods thereof. The ampicillin antibody detection kit mainly comprises an ampicillin antibody detection reagent card, ampicillin-treated erythrocyte, non-ampicillin-treated erythrocyte, ampicillin antibody positive control solution and ampicillin antibody negative control solution. The kit has the advantages ofsimplicity for operation and high sensitivity and stability, is more convenient in clinical application and can carry out rapid detection of an ampicillin antibody on 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 ampicillin 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): G01N33/577G01N33/558G01N33/559
CPCG01N33/558G01N33/559G01N33/577
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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