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Testing device for quantitatively detecting soluble growth stimulation expression protein 2

A growth stimulation and quantitative detection technology, applied in measurement devices, biological tests, material inspection products, etc., can solve the problems of inability to use heart failure prognosis assessment and severity assessment, unfavorable clinical use, slow detection speed, etc., to achieve convenient access. , to achieve the effect of accurate judgment and accurate reading

Active Publication Date: 2015-07-15
石家庄洹众生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, the enzyme-linked immunosorbent assay is used to detect growth-stimulated expression protein 2 in foreign countries. The detection speed is slow and takes about 4 hours. It is not used in clinical applications.
The patent of CN201220401468 is essentially a qualitative detection product, which cannot be used for the prognosis and severity assessment of heart failure

Method used

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  • Testing device for quantitatively detecting soluble growth stimulation expression protein 2
  • Testing device for quantitatively detecting soluble growth stimulation expression protein 2
  • Testing device for quantitatively detecting soluble growth stimulation expression protein 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: the preparation of test card of the present invention

[0046] Step 1: Preparation of anti-human ST2 monoclonal antibody

[0047]For the first basic immunization, after emulsification with Freund's complete adjuvant and equal volume of soluble ST2, 5 female BALB / c mice aged 5-7 weeks were injected at multiple points on the back of the neck. For the second immunization two weeks later, the same dose of antigen was emulsified with Freund's incomplete adjuvant and injected at multiple points on the back of the neck. Two weeks later, the titer of mouse serum was measured by enzyme-linked immunosorbent assay (ELISA); the best immunized mouse was selected, and 100 μg of antigen solution was intraperitoneally injected for booster immunization three days before fusion. The myeloma cells in the logarithmic growth phase and immune spleen cells were taken and fused with 50% PEG according to a conventional method. Soluble ST2 was coated on the microtiter plate, and ...

Embodiment 2

[0071] Embodiment 2: the preparation of test chip

[0072] The soluble ST2 is configured into a series of standard solutions such as 3.12 ng / ml, 6.25ng / ml, 12.5ng / ml, 25ng / ml, 50ng / ml, 100ng / ml, and 200ng / ml. Detect with the test card prepared in Example 1, repeat 3 times for each concentration, and read with the immunoquantitative analyzer disclosed in CN2013200892230, so as to obtain the standard curve of ST2 optical density and concentration. Due to the different reading areas of the detection line selected by different instruments, whether to filter the background or not, and the methods of calculating the optical density are different, the readings may be different. Due to differences in actual production and components of each batch of test cards, it is necessary to use this batch of test cards to analyze and obtain the standard curve.

[0073] Soluble ST2 was configured as a 3ng / ml standard solution, and the test card of Example 1 was selected for measurement, and 14 r...

Embodiment 3

[0078] Example 3 Prognosis of patients with heart failure Use the ST2 values ​​detected in Examples 1 and 2 and quantitative analysis.

[0079] Using the growth-stimulating expression protein 2 test card developed according to the above method, a total of 220 samples of patients with suspected heart failure were detected, females accounted for 64.5%, males accounted for 35.5%, and the average age was 65±16.3 years old. Growth-stimulating expression protein 2 detection kit The area under the curve of the diagnosis result is 0.919 (such as Figure 5 shown), indicating that the growth-stimulating expressed protein 2 test card has a high clinical diagnostic value and is helpful for the diagnosis and prognosis evaluation of heart failure patients. The qualitative test card cannot give data for the assessment of the severity of heart failure, and it is difficult to accurately judge due to the gray area near the critical value, let alone evaluate whether the patient has improved afte...

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Abstract

The invention relates to a testing device for quantitatively detecting a soluble growth stimulation expression protein 2. The testing device comprises a testing card and a quantitative immunity analyzer, wherein the testing card comprises a blood filtering pad, a colloidal gold pad, a nitrocellulose membrane, a water absorption pad, a back liner and a clamping shell, antihuman growth stimulation expression protein 2 antibodies marked by colloidal gold are fixed on the colloidal gold pad, detection lines coated by the antihuman growth stimulation expression protein 2 antibodies and quality control lines coated by IgG (immunoglobulin G) antibodies are arranged on the nitrocellulose membrane, a sample loading region and a detection region are arranged on the clamping shell, and the position of the detection region corresponds to the positions of the detection lines and the quality control lines; the quantitative immunity analyzer is used for detecting the optical density of the detection region of the testing card. The testing device is simple and convenient to operate and is used beside a bed, and the detection range is widened to reach 200 ng / ml maximally, so that the estimation of the prognosis and orders of severity of heart failures of different degrees is strengthened.

Description

technical field [0001] The invention relates to a test device for quantitative detection of soluble growth stimulating expression protein 2. [0002] The invention also relates to a method for quantitatively detecting soluble growth stimulating expression protein 2. Background technique [0003] Heart failure is a complex clinical syndrome with high morbidity and mortality. Clinically, the symptoms and signs of heart failure are poorly specific, and it is difficult to distinguish whether a patient is suffering from heart failure. Auxiliary examinations such as echocardiography, X-ray, and invasive hemodynamics have limitations and are limited by objective conditions. Therefore, we need to seek an objective, reliable, convenient, inexpensive, easy-to-follow-up and suitable method for dynamic observation as a supplementary method. The detection of heart failure markers can meet the above requirements. [0004] At present, there are three main markers of heart failure: B-typ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/54366G01N33/558G01N2800/325G01N2800/52
Inventor 刘成山段志强吴萌
Owner 石家庄洹众生物科技有限公司
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