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Test strip for combined and quantitative detection on urine microalbumin and cystatin C

A microalbumin and quantitative detection technology, applied in the field of disease diagnosis, can solve the problems of inability to achieve rapid bedside detection, the operation is not simple and fast enough, and the level requirements are high, and achieves rapid bedside detection, low requirements for equipment and personnel, low cost effect

Inactive Publication Date: 2018-11-30
河南省生物工程技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent document 201610227990.1 proposed "a high-performance human urine microalbumin detection kit", and 201320022738.9 proposed "a test strip for detecting cystatin C". There is no equipment for albumin or cystatin C, but there is no equipment for the joint detection of the two, and if the joint detection of urinary microalbumin and cystatin C can be realized, it will not only be easy to operate but also increase the detection rate of renal function damage. It can effectively provide an important reference for early renal function damage caused by clinical hypertension, diabetes, etc.
However, they have high requirements for laboratories and operators, and are expensive, difficult to carry out in primary hospitals, and the operation is not simple and fast enough to achieve rapid bedside detection

Method used

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  • Test strip for combined and quantitative detection on urine microalbumin and cystatin C
  • Test strip for combined and quantitative detection on urine microalbumin and cystatin C
  • Test strip for combined and quantitative detection on urine microalbumin and cystatin C

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Preparation of a test strip for combined quantitative detection of urine microalbumin and cystatin C

[0046] (1) Preparation of anti-human albumin monoclonal antibody-fluorescent microsphere complex

[0047] Place the fluorescent microspheres in 0.05M boric acid buffer for coupling, and add 0.01M EDC and 0.02M NHS buffer for activation; wash the activated fluorescent microspheres to remove EDC and NHS, add 0.05M boric acid buffer Solution, then add anti-human albumin monoclonal antibody, react at 25~30°C for 2 hours, centrifuge to obtain a precipitate; resuspend the precipitate in protective solution, block with blocking solution for 1 hour, and obtain.

[0048] (2) Preparation of cystatin C monoclonal antibody-fluorescent microsphere complex

[0049] Put the fluorescent microspheres in 0.05M boric acid buffer for coupling, and add 0.01M EDC and 0.02M NHS buffer for activation; wash the activated fluorescent microspheres to remove EDC and NHS, add 0.05M boric acid buf...

Embodiment 2

[0061] Example 2 Sample detection

[0062] (1) Quantitative detection of human urine microalbumin concentration

[0063] Use the same batch of test strips prepared in Example 1 to detect the albumin standard solution of each concentration, take the concentration of the reference product (mg / L) as the abscissa, and the fluorescence signal C / T value as the ordinate, and draw a standard curve, such as figure 2 As shown, the linear range is 10-300mg / L, and the fitting obtained standard curve equation is y=0.0185x+0.6367, R2=0.9903.

[0064] (2) Quantitative detection of cystatin C concentration in human urine

[0065] Use the same batch of test strips prepared in Example 1 to detect the cystatin C standard solution of each concentration, take the reference substance concentration (mg / L) as the abscissa, and the fluorescent signal C / T value as the ordinate, and draw a standard curve ,Such as image 3 As shown, the linear range is 0.1-8mg / L, and the standard curve equation obtai...

Embodiment 3

[0069] The performance detection of precision is carried out to the test strip prepared by the present invention.

[0070] Using the test strip prepared in Example 1, the samples with high, middle and low contents were tested continuously for 10 times, and the coefficient of variation (CV) was calculated.

[0071] The higher value of mAlb content is selected as (244.3mg / l), the middle value is selected as (86.3mg / l), and the low value is selected as (15.0mg / l), the above three urine samples are measured 10 times respectively, using CV=SD / X (SD represents the standard deviation, X represents the mean), the high value CV= 9.31%, the median CV= 8.22%, and the low value CV=7.21%.

[0072] For the higher value of Cys C content (7.13mg / l), the middle value (4.11mg / l), and the lower value (0.26ug / l), the above three urine samples were measured 10 times , using CV=SD / X, the high value CV=7.22%, the median CV=8.42%, and the low value CV=6.14%.

[0073] The CV values ​​of the detectio...

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Abstract

The invention discloses a test strip for combined quantitative detection of urine microalbumin and cystatin C, comprising a bottom plate, a sample pad, a binding pad, a nitrocellulose membrane and anabsorbent pad, wherein the bonding pad is fixedly provided with an anti-human-albumin monoclonal antibody-fluorescent microsphere complex, a cystatin C paired monoclonal antibody-fluorescent microsphere complex and a goat anti-rabbit IgG-fluorescent microsphere complex; the anti-human albumin monoclonal antibody-fluorescent microsphere complex is formed by an anti-human albumin monoclonal antibodyand fluorescent microspheres coupled by an EDC and NHS two-step method, the cystatin C monoclonal antibody-fluorescent microsphere complex is obtained by a cystatin C monoclonal antibody and fluorescent microspheres coupled by the EDC and NHS two-step method, and the goat anti-rabbit IgG-fluorescent microsphere complex is formed by sheep anti-rabbit IgG and fluorescent microspheres coupled by anEDC one-step method. The invention provides the test strip for combined quantitative detection of the urine microalbumin and cystatin C, which has the advantages of simple and convenient operation, rapid detection, high sensitivity, low cost, and suitability for point of care testing.

Description

technical field [0001] The invention relates to the technical field of disease diagnosis, in particular to a test strip for quantitatively detecting urine microalbumin and cystatin C. Background technique [0002] The physiological function of the kidney is mainly to excrete metabolic waste, regulate the acid-base balance of water and electrolytes, secrete various active substances, maintain the stability of the internal environment of the body, and ensure the normal physiological functions of the body. In recent years, due to changes in living habits and dietary structure, the incidence of hypertension and diabetes has gradually increased. And these diseases are easy to involve the kidney, leading to renal function damage, but due to the strong compensatory ability of the kidney, there are no obvious clinical features in the early stage, and are often ignored by people. When obvious symptoms appear, it is already in the middle and late stage. It has a great impact on the t...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/68G01N33/533G01N33/558
CPCG01N33/533G01N33/558G01N33/577G01N33/6893G01N2333/47G01N2800/347
Inventor 王云龙银芳颖李玉林王继创程蕾王敏
Owner 河南省生物工程技术研究中心
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