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Kit for joint detection of C-reactive protein (CRP), procalcitonin (PCT) and serum amyloid A (SAA), and preparation method of kit

A combined detection and kit technology, applied in the field of biomedicine, to achieve the effect of high sensitivity and high specificity

Inactive Publication Date: 2018-08-24
浙江艾明德生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention provides a kit for joint detection of CRP, PCT and SAA and a preparation method thereof, which overcomes the disadvantages of joint detection reagents in the prior art that cannot simultaneously detect the above three indicators rapidly and quantitatively and accurately

Method used

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  • Kit for joint detection of C-reactive protein (CRP), procalcitonin (PCT) and serum amyloid A (SAA), and preparation method of kit
  • Kit for joint detection of C-reactive protein (CRP), procalcitonin (PCT) and serum amyloid A (SAA), and preparation method of kit
  • Kit for joint detection of C-reactive protein (CRP), procalcitonin (PCT) and serum amyloid A (SAA), and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1 Preparation of coating film:

[0048] Coating buffer preparation: 0.05M pH 7.4 phosphate buffer (PB) was used as the coating buffer, which was sterilized by 0.22 μm microporous membrane and stored at 4°C for later use.

[0049] Preparation of blocking solution: 0.05M phosphate buffered saline (PBS) containing 1.0% bovine serum albumin (BSA) in mass ratio, pH 7.4, filtered through a 0.22 μm microporous membrane and stored at 4°C for later use.

[0050] Preparation of coating membrane: Dilute C-reactive protein / procalcitonin / serum amyloid A antibody to 0.5mg / ml with 0.05M phosphate buffer (PB), pH7.4 coating buffer, goat anti-mouse The secondary antibody was diluted to 1 mg / ml, and was uniformly spray-printed at 0.5 cm intervals on NC membranes with a width of 3.5 cm using a quantitative spraying device at 1 μl / cm. 0.05M PBS, pH 7.4) for 10 minutes, dried at 25-35°C for 8 hours, added desiccant and sealed for later use.

Embodiment 2

[0051] Example 2 Preparation of magnetic particle-labeled antibody:

[0052] Preparation of citric acid buffer: Weigh 14.7g of sodium citrate and 10.5g of citric acid and dissolve in purified water to prepare 1L of citric acid buffer with a pH value of 4.6 and a concentration of 0.05M, and add Tween-20 with a volume ratio of 0.05%. Filter and sterilize with a 0.22 μm microporous membrane and store at 4°C for later use.

[0053] Preparation of boric acid storage buffer: Weigh 0.62g of boric acid and 3.81g of borax and dissolve in purified water to prepare 1L of boric acid buffer solution with a pH of 9.0 and a final concentration of 0.05M, and add povidone (PVP) with a mass ratio of 1%. 0.5% Casein, 0.5% Tween-20 by volume, 5% sucrose by mass, sterilized by 0.22 μm microporous membrane and stored at 4°C for later use.

[0054] Preparation of magnetic particles:

[0055] 1. Preparation of magnetic particles containing CRP antibody:

[0056] (1) Wash the magnetic particles wit...

Embodiment 3

[0071] Example 3 Assembly of test strips and finished products

[0072] All of the following operating environment requirements are: humidity less than 20%, temperature 18-26 ℃.

[0073] Assembling the test board: use the BioDot LM5000 assembly machine to assemble the 3.5cm coating film on the transparent plastic base plate with a width of 9.8cm according to the requirements, cover the upper transparent plastic cover, and assemble the test board.

[0074] Cutting of test strips: Use a BioDot CM4000 strip cutter to cut the assembled test strips into 0.5cm wide test strips, then pack them into 20 strips / tube, and store them at room temperature.

[0075] D. Aliquoting of magnetic particle-labeled antibody

[0076] The prepared magnetic particles containing the three antibodies were divided into 25mL / bottles and stored at 2-8°C.

[0077] E. Finished product packaging

[0078] A tube filled with 20 test strips and a glass bottle filled with 25mL mixed magnetic particle-labeled a...

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Abstract

The invention provides a kit for joint detection of C-reactive protein (CRP), procalcitonin (PCT) and serum amyloid A (SAA), and a preparation method of the kit. The kit comprises a CRP monoclonal antibody solution, a PCT monoclonal antibody solution and an SAA monoclonal antibody solution which are respectively labeled with magnetic particles, a carrier enveloped with a CRP monoclonal antibody, acarrier enveloped with a PCT monoclonal antibody, a carrier enveloped with an SAA monoclonal antibody, a carrier enveloped with a goat anti-mouse antibody, and a nitrocellulose membrane test strip. The preparation method comprises the following steps: respectively labeling the CRP monoclonal antibody, the PCT monoclonal antibody and the SAA monoclonal antibody with the magnetic particles; respectively enveloping the carriers with the CRP monoclonal antibody, the PCT monoclonal antibody, the SAA monoclonal antibody and the goat anti-mouse antibody; subpackaging and assembling to obtain the finished product. The kit can rapidly, quantitatively and accurately detect three markers at the same time.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a kit for joint detection of CRP, PCT and SAA and a preparation method thereof. Background technique [0002] C-reactive protein (C-neactveprotein, referred to as CRP), discovered as early as 1930, is an acute phase reaction protein that can react with pneumococcal C polysaccharide to form a complex. And when the body is infected or tissue damage, some proteins (acute proteins) that rise sharply in the plasma activate complement and strengthen the phagocytosis of phagocytic cells to play a conditioning role, and remove pathogenic microorganisms that invade the body and damaged, necrotic, and apoptotic tissue cells . In recent years, due to the update of detection technology, a fast, simple and reliable method for measuring CRP has been rapidly established. The clinical application of CRP has greatly increased, and its medical value is being widely verified and recognized. CRP rises ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/543
CPCG01N33/54326G01N33/558G01N33/577
Inventor 胡国富朱炎
Owner 浙江艾明德生物科技有限公司
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