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Time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and preparation method thereof

A time-resolved fluorescence and quantitative detection technology, applied in the field of clinical medical diagnosis, can solve problems such as background signal interference, unsatisfactory sensitivity, long detection time, etc., to improve detection precision and accuracy, reduce background signal value, and exclude samples the effect of interference

Inactive Publication Date: 2016-12-21
SHANGHAI UPPER BIO TECH PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Enzyme-linked immunosorbent assay is complex in operation, long in detection time, and narrow in detection range; latex-enhanced immunoturbidimetry needs to be combined with large-scale instruments, and the cost is high; colloidal gold immunochromatography for qualitative or quantitative detection has low sensitivity and narrow linear range, which cannot meet quantitative , Accurate detection requirements; Most of the fluorescent immunochromatography methods use fluorescein or other substances as light sources. Although they have a high linear range, they cannot guarantee low-end sensitivity due to background signal interference.
[0006] Although time-resolved fluorescent immunochromatography has been widely used in the detection of various items, most of them use time-resolved fluorescent microspheres immobilized on the conjugate release pad, and the imprecision is large due to the uneven release of the microspheres. , can’t meet the items with high sensitivity requirements, especially the detection of troponin I
In addition, in order to detect whole blood samples, the sample pads of most manufacturers use artificially treated glass fibers as conjugate release pads, which also increases the imprecision

Method used

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  • Time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and preparation method thereof
  • Time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and preparation method thereof
  • Time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] 1. Preparation of CK-MB time-resolved fluorescent immunochromatographic test strips

[0054] (1) Preparation of test line (T) solution: Dilute CK-MB monoclonal antibody 1 to 0.5 mg / ml with 50 mmol / L citrate solution;

[0055] (2) Preparation of quality control line (C) solution: dilute rabbit IgG antibody to 0.5mg / ml with 50mmol / L citrate solution;

[0056] On the bottom plate (1) with adhesive backing, adopt the overlapping method, first paste the nitrocellulose membrane (3), and then paste the Fusion5 membrane (2) and the absorbent paper ( 4). On the nitrocellulose membrane (3) and close to the Fusion5 membrane (2), draw T (CK-MB capture line (5)), draw C (rabbit IgG) line on the end close to the absorbent paper (4), T, C The spacing is 5mm.

[0057] Draw a T line at the T line and a C line at the C line. The final concentration of T-line antibody is 0.5mg / ml, the final concentration of C-line rabbit IgG is 0.5mg / ml, and the spray volume of C and T-line is 0.5μl / c...

Embodiment 2

[0081] 1. Preparation of CK-MB time-resolved fluorescent immunochromatographic test strips

[0082] (1) Preparation of test line (T) solution: Dilute CK-MB monoclonal antibody 1 to 1 mg / ml with 50 mmol / L citrate solution;

[0083] (2) Preparation of quality control line (C) solution: dilute rabbit IgG antibody to 1 mg / ml with 50 mmol / L citrate solution;

[0084]On the bottom plate (1) with adhesive backing, adopt the overlapping method, first paste the nitrocellulose membrane (3), and then paste the Fusion5 membrane (2) and the absorbent paper ( 4). On the nitrocellulose membrane (3) and close to the Fusion5 membrane (2), draw T (CK-MB capture line (5)), draw C (rabbit IgG) line on the end close to the absorbent paper (4), T, C The spacing is 5mm.

[0085] Draw a T line at the T line and a C line at the C line. The final concentration of T-line antibody is 1 mg / ml, the final concentration of C-line rabbit IgG is 1 mg / ml, and the spray volume of C and T lines is 1.0 μl / cm. ...

Embodiment 3

[0108] 1. Preparation of CK-MB time-resolved fluorescent immunochromatographic test strips

[0109] (1) Preparation of test line (T) solution: Dilute CK-MB monoclonal antibody 1 to 2 mg / ml with 50 mmol / L citrate solution;

[0110] (2) Preparation of quality control line (C) solution: dilute rabbit IgG antibody to 2mg / ml with 50mmol / L citrate solution;

[0111] On the bottom plate (1) with adhesive backing, adopt the overlapping method, first paste the nitrocellulose membrane (3), and then paste the Fusion5 membrane (2) and the absorbent paper ( 4). On the nitrocellulose membrane (3) and close to the Fusion5 membrane (2), draw T (CK-MB capture line (5)), draw C (rabbit IgG) line on the end close to the absorbent paper (4), T, C The spacing is 5mm.

[0112] Draw a T line at the T line and a C line at the C line. The final concentration of T-line antibody is 2 mg / ml, the final concentration of C-line rabbit IgG is 2 mg / ml, and the spray volume of C and T lines is 2.0 μl / cm. ...

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Abstract

The invention discloses a time-resolved fluorescence immunochromatography reagent used for rapid quantitative detection of CK-MB, and a preparation method thereof, and belongs to the field of clinical diagnose. The time-resolved fluorescence immunochromatography reagent is composed of a test paper strip and a fluorescent liquid; the test paper strip comprises a base plate, Fusion5, a nitrocellulose membrane, and a water absorption pad; Fusion5, the nitrocellulose membrane, and the water absorption pad are connected in sequence in the horizontal direction, and are fixedly arranged on the base plate; the nitrocellulose membrane is coated with a CK-MB monoclonal antibody 1 detection line and a quality control line composed of rabbit IgG antibodies; and the fluorescent liquid comprises CK-MB monoclonal antibody 2 labelled fluorescent microspheres, and goat anti-rabbit antibody labeled fluorescent microspheres. According to the time-resolved fluorescence immunochromatography reagent, time-resolved fluorescent microspheres are used for increasing fluorescence intensity, reducing background signals, and realizing quantitative detection of CK-MB content of whole blood, serum, or blood plasma at the same time; and only 10 to 20ml of a sample is needed. Application of the time-resolved fluorescence immunochromatography reagent is convenient and rapid; operation is simple; detection time is short; specificity and sensitivity are high; detection results are more accurate; and the time-resolved fluorescence immunochromatography reagent is suitable for clinical POCT rapid diagnosis.

Description

technical field [0001] The invention belongs to the field of clinical medical diagnosis, and in particular relates to a time-resolved fluorescence immunochromatography reagent for rapid and quantitative detection of CK-MB and a preparation method thereof. Background technique [0002] CK-MB is one of the three isoforms of creatine kinase (CK) (the other two are CK-BB and CK-MMOL / L). CK is a main enzyme of muscle metabolism. CK-MB consists of two subunits (MW=40000 each): subunit M for muscle and subunit B for brain. CK-MB mainly exists in the myocardium, and a small amount exists in the skeletal muscle. Quantitative determination of CK-MB is part of routine cardiac examination and is helpful for the diagnosis of AMI. When AMI occurs, CK-MB appears in the blood circulation and indicates that the myocardium has been damaged. CK-MB concentrations rise within 4-8 hours of pain onset, peak at 12-24 hours, and then decline to normal levels after 48 hours. CK-MB is the marker ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/533G01N33/543G01N21/64
CPCG01N33/558G01N21/6408G01N33/533G01N33/54313
Inventor 朱传增石晓强安永君李福刚徐建新
Owner SHANGHAI UPPER BIO TECH PHARMA
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