Test strip and kit for detecting mycophenolic acid, and preparation method of test strip

A technology of mycophenolic acid and test strips, which is applied in the field of mycophenolic acid detection, can solve the problems of inability to achieve monitoring and cannot be made into single-serve packaging, and achieve the effects of easy operation, wide range of detection sample types, and good stability

Active Publication Date: 2019-10-25
BEIJING DIAGREAT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method requires the use of a fully automatic biochemical analyzer, and the storage temperature is 2-8°C. It needs to be transported in a cold chain. It cannot be packaged as a single serving, and it cannot achieve the purpose of timely monitoring in the drug department.

Method used

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  • Test strip and kit for detecting mycophenolic acid, and preparation method of test strip
  • Test strip and kit for detecting mycophenolic acid, and preparation method of test strip
  • Test strip and kit for detecting mycophenolic acid, and preparation method of test strip

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Effect test

preparation example Construction

[0033] The present invention also provides a preparation method for the test strip, comprising the following steps: 1) treating the sample pad with a sample pad treatment solution to obtain a treated sample pad; 2) mixing the mycophenolic acid monoclonal antibody with the conjugate After mixing and coupling, block to obtain mycophenolic acid specific antibody conjugates; spray the mycophenolic acid specific antibody conjugates on the treated glass fiber membrane to obtain coated mycophenolic acid specific antibody conjugates 3) spray mycophenolic acid protein conjugates on the detection line of the nitrocellulose membrane, and spray goat anti-mouse IgG on the quality control line; obtain the nitrocellulose sprayed with detection line and quality control line Membrane; 4) The treated sample pad, the glass cellulose membrane coated with the mycophenolic acid specific antibody conjugate, the nitrocellulose membrane provided with the detection line and the quality control line, and...

Embodiment 1

[0043] Preparation of test strips:

[0044] 1) Soak the sample pad in the sample pad treatment solution for 0.5h, and dry in an oven at 65°C for 2h;

[0045] 2) Use fluorescent microspheres with a particle size of 100nm; add 1% fluorescent microspheres, EDC 10mg / mL, mycophenolate monoclonal antibody 200μg / mL, solvent 20mmol / L phosphate buffer, mix and couple for 3h, Centrifuge to remove the supernatant, then add 1% BSA to block for 1h.

[0046] 3) Centrifuge the prepared labeled conjugate and use resuspension buffer (50mm TRIS buffer, polyethylene glycol 3g / L, alum 1.2g / L, mannitol 4g / L, sodium chloride 0.9%) Resuspend the labeled conjugate to a concentration of 0.2%, soak the glass cellulose membrane in pretreatment buffer (the composition content is consistent with the reselection buffer) for 2 hours, and dry for 1 hour; Spray the film onto the glass cellulose film. After the gold spraying is completed, dry it in a blast oven at 65°C for 2 hours.

[0047] 4) The T line is...

Embodiment 2

[0057] Preparation of test strips:

[0058] 1) Take 0.01% HAuCl 4 100mL aqueous solution, heated to boil, quickly added 0.75mL 1% trisodium citrate aqueous solution, continued to boil for about 5min, orange-red color appeared, and 50nm colloidal gold particles were produced.

[0059] 2) Colloidal gold solution, mycophenolate monoclonal antibody 200 μg / mL, mixed and coupled for 3 hours, centrifuged to remove the supernatant, and then added 1% BSA to block for 1 hour.

[0060] 3) Centrifuge the prepared labeled conjugate and use resuspension buffer (20mm borate buffer, polyethylene glycol 0.5g / L, alum 0.2g / L, mannitol 4g / L, sodium chloride 0.9%) For resuspension, the gold pad was soaked in pretreatment buffer (consisting with the resuspension buffer) for 2 hours, and then dried for 1 hour; using a gold spraying device, spray the resuspension onto the gold pad, and after the gold spraying was completed 45 ℃ Blast drying oven, drying 6h;

[0061]4) The T line on the nitrocellul...

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Abstract

The invention provides a test strip and a kit for detecting mycophenolic acid, and a preparation method of the test strip, which belong to the technical field of mycophenolic acid detection. The teststrip comprises a bottom plate, and a sample pad, a glass fiber membrane, a nitrocellulose membrane and water absorbing paper which are sequentially in overlap joint with the surface of the bottom plate, wherein the sample pad is treated with a sample pad treatment liquid; the glass fiber membrane is treated with a glass fiber membrane treatment liquid; the glass fiber membrane is coated with a mycophenolic acid specific antibody conjugate; and a detection line and a quality control line are arranged on the nitrocellulose membrane, and a mycophenolic acid protein conjugate is sprayed on the detection line. The test strip provided by the invention can quickly detect the content of mycophenolic acid in a blood sample, and has the advantages of good stability, wide linearity range, low detection cost and simple operation.

Description

technical field [0001] The invention belongs to the technical field of mycophenolic acid detection, in particular to a test strip for detecting mycophenolic acid, a kit and a preparation method of the test strip. Background technique [0002] Mycophenolic acid (MPA), also known as mycophenolic acid, can non-competitively bind hypoxanthine mononucleotide dehydrogenase, which is de novo synthesis of guanine nucleotides during the proliferation of T and B lymphocytes key enzymes. As the main immunosuppressant, mycophenolate mofetil derivatives have been widely used at home and abroad to prevent and treat acute rejection of transplanted organs. Mycophenolate mofetil exerts immunosuppressive activity in vivo through conversion to mycophenolic acid. In different transplant populations, great differences in the pharmacokinetics of mycophenolic acid can be found among individuals, and internal and external factors can affect its pharmacokinetics. [0003] With the initiative of p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N33/9493G01N33/558G01N33/54366G01N2430/00G01N33/54388
Inventor 周建平周裕军许秀丽
Owner BEIJING DIAGREAT BIOTECH CO LTD
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