Detection kit and detection method of serum amyloid A protein

A detection kit and serum starch technology, applied in the field of immune analysis, can solve the problems of large blood volume and inability to detect, and achieve good sensitivity and precision

Inactive Publication Date: 2016-09-07
SHENZHEN GOLDSITE DIAGNOSTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of nephelometry for SAA, the kits in the prior art can only detect SAA in serum. Although they can resist the interference of certain hemoglobin, the detection cannot be performed if the hemoglobin content is higher than 4g / L.
The content of hemoglobin in normal human blood is 110g / L~160g / L, therefore, the kits in the prior art cannot realize the detection of whole blood
However, the amount of blood required for venous blood separation and serum separation is large

Method used

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  • Detection kit and detection method of serum amyloid A protein
  • Detection kit and detection method of serum amyloid A protein
  • Detection kit and detection method of serum amyloid A protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] Embodiment 1 Kit provided by the invention

[0113] Reaction solution components:

[0114]

[0115] SAA antibody suspension components:

[0116]

[0117] Calibrator solution components:

[0118]

[0119] Wherein, the reaction solution is prepared by mixing the components, filtering through a 0.22-micron membrane filter, subpackaging, and storing at -20°C.

[0120] Antibody suspensions were prepared as follows:

[0121]Take 10 mg of carboxypolystyrene microspheres, add an appropriate amount of 50 mM MES pH6.0 buffer to wash 2-3 times, and finally use 50 mM MES pH6.0 buffer to dilute to 0.5% (mass volume ratio) concentration. Add an appropriate amount of carbodiimide and N-hydroxysulfosuccinimide to the activated polystyrene microspheres, stir at room temperature for 20 min, wash twice with 20mM HEPES pH7.5 buffer, and set the temperature with 20mM HEPES pH7.5 buffer. Contain to 1% (mass volume ratio) concentration. Add an equal volume of activated and washe...

Embodiment 2

[0124] Embodiment 2 kit provided by the invention

[0125] Reaction solution components:

[0126]

[0127] SAA antibody suspension components:

[0128]

[0129] Calibrator solution components:

[0130]

[0131] Wherein, the reaction solution is prepared by mixing the components, filtering through a 0.22-micron membrane filter, subpackaging, and storing at -20°C.

[0132] Antibody suspensions were prepared as follows:

[0133] Take 10 mg of carboxypolystyrene microspheres, add an appropriate amount of 25mM MES pH6.0 buffer to wash 2-3 times, and finally use 25mM MES pH6.0 buffer to dilute to 1% (mass volume ratio) concentration. Add an appropriate amount of carbodiimide and N-hydroxysulfosuccinimide to the activated polystyrene microspheres, stir at room temperature for 20 min, wash twice with 50mM HEPES pH7.5 buffer, and fix with 20mM HEPES pH7.5 buffer. Contain to 0.5% (mass volume ratio) concentration. Add equal volumes of activated and washed microspheres to 0...

Embodiment 3

[0136] Embodiment 3 Kit provided by the invention

[0137] Reaction solution components:

[0138]

[0139] SAA antibody suspension components:

[0140]

[0141]

[0142] Calibrator solution components:

[0143]

[0144] Wherein, the reaction solution is prepared by mixing the components, filtering through a 0.22-micron membrane filter, subpackaging, and storing at -20°C.

[0145] Antibody suspensions were prepared as follows:

[0146] Take 10 mg of carboxypolystyrene microspheres, add an appropriate amount of 25mM MES pH6.0 buffer to wash 2-3 times, and finally use 25mM MES pH6.0 buffer to dilute to 0.5% (mass volume ratio) concentration. Add appropriate amount of carbodiimide and N-hydroxysulfosuccinimide to the activated polystyrene microspheres, stir at room temperature for 20min, wash twice with 25mM MES pH6. Contain to 0.5% (mass volume ratio) concentration. Add an equal volume of activated and washed microspheres to 0.05% SAA antibody solution, stir at r...

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Abstract

The invention relates to the field of immunoassay, and especially relates to a detection kit and a detection method of a serum amyloid A protein. In the kit, a reaction solution well cooperates with a whole blood sample in order to make the kit directly detect the whole blood, so the kit can complete detection only through finger tip blood without drawing venous blood or separating serum. Compared with kits in the prior art, the kit provided by the invention has the advantages of wider linear range (2.4-240mg / L), lower detection limit (2.39mg / L), good linear range and sensitivity, average recovery rate of the detection sample of 101.87%, proportional systematic error being smaller than 5%, and good accuracy. The variation coefficient is smaller than 3% when the kit is used to respectively carry out high-value quality control and low-value quality control 10 times, so the kit has good precision.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a detection kit and a detection method for serum amyloid A. Background technique [0002] Serum amyloid A (serum amyloid A protein, SAA) is an acute phase reaction protein. During acute inflammatory reactions such as infection and trauma, the concentration of SAA rises very rapidly, and it is stimulated by IL-1, IL-6 and TNF in a short time. Stimulation, which is synthesized in large quantities by activated macrophages and fibroblasts in the liver, can increase to more than 1000 times the initial concentration, and this characteristic also makes SAA one of the most sensitive markers of acute inflammation at present. At the same time, in various chronic inflammatory pathological states including cardiovascular disease, obesity, and ankylosing spondylitis, extrahepatic tissues such as human adipocytes, vascular endothelial cells, and monocyte-macrophages can also synthesize and secre...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N33/68G01N33/96G01N21/47
CPCG01N33/68G01N21/47G01N33/96
Inventor张宁陈明峰
OwnerSHENZHEN GOLDSITE DIAGNOSTICS