Kit for detecting blood cortisol of animals

A technology for detecting animals and cortisol, applied in the field of immunoassay, can solve the problems of high requirements for experimental skills, many manual operation steps, and complicated operation steps, and achieves the effects of low requirements for experimental skills, small amount of reagents, and simple and convenient operation.

Pending Publication Date: 2019-03-08
CHENGDU POLYTECH BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of enzyme-linked immunosorbent assay to determine the blood cortisol content of dogs, cats and other animals has the following disadvantages: the method is not sensitive enough, suitable for qualitative and semi-quantitative determination, and the operation steps are cumbersome, and many reagents need to be configured by the customer. The experimental skills required are high, and the amount of reagents used is large. Due to many manual steps, errors are prone to occur.
[0008] 1. It can only be used for the determination of cortisol content in human blood, but not for the determination of cortisol content in animal blood;
[0009] 2. When using this kit to measure the cortisol content in blood, there are still cumbersome operation steps, and many reagents need to be configured by the customer. The experimental skills of the experimental operators are high, and the amount of reagents is large, which is prone to errors.

Method used

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  • Kit for detecting blood cortisol of animals
  • Kit for detecting blood cortisol of animals
  • Kit for detecting blood cortisol of animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] 1. Preparation of Cortisol Monoclonal Antibody Coated Capillary

[0073] Cortisol monoclonal antibody-coated capillaries are specifically:

[0074] The high-precision capillary tube is selected for the capillary, the outer diameter is 1.18±0.02mm, the inner diameter is 0.7±0.005m, and the length is 30±1mm; the material is borosilicate 3.3 glass.

[0075] Cortisol monoclonal antibody-coated capillaries are made by the following methods:

[0076] Step 1, evenly wrap the amino group on the inner surface of the capillary

[0077] Step 2. Coating the capillary

[0078] First, dissolve EDC with MES buffer to make solution I; then dilute the cortisol monoclonal antibody with solution I to make a coating solution; finally, soak the capillary after step 1 with the coating solution at 37°C. Soaking time is 40min, pour out the coating liquid after soaking;

[0079] The pH of the MES buffer is 6.4, and the concentration is 0.1M; the EDC concentration in the solution I is 0.1 mg / mL; the concen...

Embodiment 2

[0108] This embodiment is basically the same as embodiment 1, the difference is: MES buffer includes the following components: MES: 1.5M, NaCl: 1.5M, MgCl 2 :1mM, ZnCl 2 : 0.01mM, BSA: 0.2%, Tween 20: 0.2%;

[0109] The coating temperature is 40°C, the coating time is 1 hour, and the coating method is to immerse the capillary in the coating liquid.

[0110] The pH of the MES buffer was 6.6, and the concentration was 0.2M; the EDC concentration in solution I was 0.8 mg / mL; the concentration of the cortisol monoclonal antibody in the coating solution was 20 ug / mL.

[0111] The pH of the Tris-HCl buffer is 7.4 and the concentration is 100 mM.

[0112] The sealing treatment time of the sealing liquid is 5h, and the sealing treatment temperature is 40°C.

[0113] The blocking solution includes the following components: BSA: 5%, sucrose: 10%, Tris: 20M, glycine: 0.05M, and Tween 20: 0.01%.

[0114] The cleaning agent contains Tris, NaCl, CHAPS and Trition 100, wherein the concentration of Tris...

Embodiment 3

[0123] This embodiment is basically the same as embodiment 1, the difference is: MES buffer includes the following components: MES: 1M, NaCl: 1M, MgCl 2 : 2mM, ZnCl 2 : 0.05mM, BSA: 0.8%, Tween 20: 0.4%;

[0124] The coating temperature is 38°C, the coating time is 1 hour, and the coating method is to immerse the capillary in the coating liquid.

[0125] The pH of the MES buffer is 6.5, and the concentration is 0.1M; the concentration of EDC in solution I is 0.3 mg / mL; the concentration of the cortisol monoclonal antibody in the coating solution is 6 ug / mL.

[0126] The pH of the Tris-HCl buffer is 7.4 and the concentration is 40 mM.

[0127] The sealing treatment time of the sealing liquid is 3h, and the sealing treatment temperature is 22°C.

[0128] The blocking solution includes the following components: BSA: 3%, sucrose: 5%, Tris: 120M, glycine: 0.4M, and Tween 20: 0.06%.

[0129] The cleaning agent contains Tris, NaCl, CHAPS and Trition 100, wherein the concentration of Tris is 120...

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Abstract

The invention discloses a kit for detecting blood cortisol of animals. The kit comprises a capillary tube coated with cortisol monoclonal antibodies, an alkaline phosphatase-labeled cortisol antigen analog, a chemiluminescent substrate for catalyzing alkaline phosphatase luminescence, an ANS salt release agent, a cortisol calibrator and a cleaning solution. When being used for measuring the bloodcortisol content of the animals such as dogs and cats, the kit has the advantages of simple operation, high sensitivity, high specificity, wide linear range, high stability and small reagent dosage.

Description

Technical field [0001] The invention belongs to the technical field of immunoassays, and particularly relates to a kit for detecting cortisol in animal blood. Background technique [0002] Cortisol, also known as hydrocortisone, is an adrenal cortex hormone secreted from the adrenal cortex that has the strongest effect on carbohydrate metabolism. The secretion of cortisol is mainly regulated by the adrenocorticotropic hormone secreted by the pituitary gland. Its secretion has a clear circadian rhythm. The secretion reaches a peak around 8 am, and then gradually decreases, reaching the lowest value at midnight. Cortisol has a particularly important effect in manipulating mood and health, immune cells and inflammation, blood vessels and blood pressure, and maintaining connective tissues. [0003] Cortisol exists in the blood in two forms: bound state and free state. Cortisol in the free state only accounts for about 10% and has biological activity; the bound state is mainly combine...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N33/552G01N33/577
CPCG01N33/552G01N33/577
Inventor高翠华冉鹏
OwnerCHENGDU POLYTECH BIOLOGICAL TECH CO LTD