Immunofluorescence chromatography kit for quantitative detection of SAA (serum amyloid A), CRP (C-reactive protein) and PCT (procalcitonin) and preparation method of immunofluorescence chromatography kit

A quantitative detection and immunofluorescence technology, applied in measurement devices, analytical materials, instruments, etc., can solve the problems of difficult to meet quantitative detection requirements, poor sensitivity, low accuracy, etc., and improve the accuracy of poor accuracy, high sensitivity, and accurate results. Effect

Inactive Publication Date: 2017-09-15
深圳市惠安生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technology sets three detection lines and one detection line on a test strip at the same time, and detects three indicators simultaneously. The results are judged by the matching immunofluorescence analyzer, which has poor sensitivity and low accuracy, and it is difficult to meet the quantitative detection requirements of clinical requirements.

Method used

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  • Immunofluorescence chromatography kit for quantitative detection of SAA (serum amyloid A), CRP (C-reactive protein) and PCT (procalcitonin) and preparation method of immunofluorescence chromatography kit
  • Immunofluorescence chromatography kit for quantitative detection of SAA (serum amyloid A), CRP (C-reactive protein) and PCT (procalcitonin) and preparation method of immunofluorescence chromatography kit
  • Immunofluorescence chromatography kit for quantitative detection of SAA (serum amyloid A), CRP (C-reactive protein) and PCT (procalcitonin) and preparation method of immunofluorescence chromatography kit

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Experimental program
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preparation example Construction

[0037] Further, the present invention also provides a method for preparing the above-mentioned immunofluorescence chromatography kit for quantitative detection of SAA, CRP, and PCT, comprising the following steps:

[0038] 1) Preparation of fluorescent immunochromatography test strips: paste nitrocellulose membrane on the backing of the test paper, affix an absorbent pad on one end of the nitrocellulose membrane, affix a sample pad on the other end, and absorb it near the nitrocellulose membrane. One end of the pad is sequentially marked with goat anti-rabbit IgG polyclonal antibody quality control line, SAA monoclonal antibody detection line, CRP monoclonal antibody detection line and PCT monoclonal antibody detection line to prepare a test paper board, and then use a strip cutter to cut the test paper The plate is cut longitudinally into 4mm wide test strips;

[0039] 2) Preparation of the fluorescent substance application solution: respectively couple the rabbit IgG polyclo...

Embodiment 1

[0045] This embodiment provides an immunofluorescence chromatography kit for the quantitative detection of SAA, CRP, and PCT and a preparation method thereof.

[0046] In this embodiment, the fluorescent substance is a time-resolved fluorescent microsphere with an excitation wavelength of 360 nm and an emission wavelength of 615 nm, which is a fluorescent dye polystyrene microsphere coated with rare earth europium element.

[0047] The preparation method of the immunofluorescence chromatography kit for the quantitative detection of SAA, CRP, PCT of this embodiment comprises the following steps:

[0048] (1) Activation of time-resolved fluorescent microspheres: First, take 1 mL of fluorescent microspheres and add 4 ml of 0.05 mol / L, pH 6.0 MES solution, and ultrasonically disperse them; weigh 5 mg of N-hydroxysuccinimide (NHS) and add into the microsphere suspension, then shake well to dissolve; then weigh 5 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC...

Embodiment 2

[0057] This embodiment provides an immunofluorescence chromatography kit for the quantitative detection of SAA, CRP, and PCT and a preparation method thereof.

[0058] The structure of the test strip in this example is the same as that in Example 1, the difference is that the fluorescent substance in Example 2 is a fluorescent microsphere with an excitation wavelength of 470nm-490mm and an emission wavelength of 525-560nm. Dyed polystyrene microspheres.

[0059] The preparation method of the immunofluorescence chromatography kit for the quantitative detection of SAA, CRP, PCT of this embodiment comprises the following steps:

[0060] (1) Activation of fluorescent microspheres: first take 1mL of fluorescent microspheres and use ultrasonic waves to disperse them by ultrasonic waves. The treatment time is 30s with 100W ultrasonic waves; min, centrifuged for 20 min, poured off the supernatant, and added 1 mL of 0.1 mol / L, pH 4.7 MES buffer. Weigh 3 mg of N-hydroxysuccinimide (NH...

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Abstract

The invention discloses an immunofluorescence chromatographic kit for quantitative detection of SAA, CRP and PCT and a preparation method thereof, comprising a fluorescent immunochromatographic test strip and a fluorescent substance. The fluorescent immunochromatographic test strip is provided with Nitrocellulose membrane, one end of the nitrocellulose membrane is connected with a sample pad, and the other end of the nitrocellulose membrane is connected with an absorption pad; the detection line for detecting the content of SAA, CRP and PCT is arranged in parallel on the nitrocellulose membrane The fluorescent substance includes a fluorescent substance coupled to a rabbit IgG polyclonal antibody, a fluorescent substance coupled to a paired SAA monoclonal antibody, a fluorescent substance coupled to a paired CRP monoclonal antibody, and a paired PCT monoclonal antibody of fluorescent substances. The kit can detect the contents of SAA, CRP and PCT simultaneously conveniently, accurately and with high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a rapid and quantitative detection reagent, which adopts fluorescent microspheres, quantum dots, etc. as markers, uses immunochromatographic methods, and uses a fluorescence immunoassay analyzer to simultaneously detect human serum and plasma and the contents of SAA, PCT and CRP in whole blood samples, specifically an immunofluorescence chromatography kit for quantitative detection of SAA, CRP and PCT and a preparation method thereof. Background technique [0002] Serum amyloid A (SAA) is an acute time-limited reaction protein, and the concentration of serum amyloid A is a sensitive indicator reflecting the early inflammation of infectious diseases, which is helpful for diagnosing inflammation, evaluating its activity, monitoring its activity and treatment . [0003] C-reactive protein (CRP) refers to an acute phase protein whose plasma content rises sharply when...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/543G01N33/533
CPCG01N33/558G01N33/533G01N33/543
Inventor 梁培华梁巍腾刘云青高翔陈志兴
Owner 深圳市惠安生物科技有限公司
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