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Enzyme linked immunosorbent assay kit for detecting medroxyprogesterone and application thereof

An enzyme-linked immunosorbent reagent and medroxyprogesterone technology, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of high cost of capital and personnel input, and achieve a simple pretreatment process, high sensitivity, and convenient use. Effect

Active Publication Date: 2014-12-17
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods for detecting such drugs are mainly gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry and other instrument methods, and the investment costs of capital and personnel are relatively high.

Method used

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  • Enzyme linked immunosorbent assay kit for detecting medroxyprogesterone and application thereof
  • Enzyme linked immunosorbent assay kit for detecting medroxyprogesterone and application thereof
  • Enzyme linked immunosorbent assay kit for detecting medroxyprogesterone and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Preparation of kit components

[0028] 1. Preparation of medroxyprogesterone hapten

[0029] The mixture of 0.70 g medroxyprogesterone in 5 ml dimethyl sulfoxide (DMSO), slowly added dropwise at 40 °C, the mixture of 0.5 ml 1,3-propanediamine and 0.5 ml pyridine in 10 ml DMSO, dropwise After completion, the reaction was continued for 12 hours, the solvent and unreacted propylenediamine were removed by rotary evaporation, and medroxyprogesterone-3-aminopropyl oxime was quantitatively obtained.

[0030] Get above-mentioned product and measure through proton nuclear magnetic resonance spectrum, such as figure 2 As shown, the increased methylene peak between 1.0-2.5 indicates that the target hapten was synthesized successfully.

[0031] 2. Antigen preparation

[0032] Immunogen preparation—the immunogen was obtained by coupling the medroxyprogesterone hapten with bovine serum albumin (BSA).

[0033] Take 30 mg of medroxyprogesterone-3-aminopropyl oxime and...

Embodiment 2

[0045] Example 2 Formation of an enzyme-linked immunosorbent assay kit for detecting medroxyprogesterone

[0046] An enzyme-linked immunosorbent assay kit for detecting medroxyprogesterone was set up to include the following components:

[0047] (1) Enzyme plate coated with medroxyprogesterone-conjugated antigen;

[0048] (2) 6 bottles of medroxyprogesterone standard solution, the concentrations are 0μg / L, 0.03μg / L, 0.06μg / L, 0.12μg / L, 0.24μg / L, 0.48μg / L;

[0049] (3) Medroxyprogesterone monoclonal antibody labeled with horseradish peroxidase;

[0050] (4) The substrate chromogenic solution is composed of liquid A and liquid B, liquid A is carbamide peroxide, and liquid B is tetramethylbenzidine;

[0051] (5) The stop solution is 2mol / L sulfuric acid;

[0052] (6) The washing solution has a pH value of 7.4, contains 0.5%~1.0% Tween-20, 0.01‰~0.03‰ sodium azide preservative, and 0.1~0.3mol / L phosphate buffer, the percentage is weight volume percentage;

[0053] (7) The com...

Embodiment 3

[0054] Example 3 Detection of medroxyprogesterone in animal tissue samples

[0055] 1. Sample pretreatment

[0056] Homogenize the tissue sample with a homogenizer; weigh 3.0±0.05g of the homogenate into a 50ml polystyrene centrifuge tube, measure 90ml of acetonitrile and add it to 10ml of 0.1mol / L sodium hydroxide solution, mix well, add 6ml of the acetonitrile -0.1mol / L sodium hydroxide solution, vortex for 2min, over 3000g, centrifuge at room temperature for 5min; take out 1ml of supernatant into a 15ml clean glass test tube, blow dry under nitrogen flow at 50-60℃; add 1ml of reconstitution working solution , use a vortex instrument to vortex for 1min; take 100ul and add 900ul reconstituted working solution, mix well; take 50ul for detection.

[0057] 2. Detection with kit

[0058] Add 50 μl of medroxyprogesterone standard solution or pretreated sample solution to the corresponding microwells of the microplate plate coated with medroxyprogesterone-coupled antigen, then ad...

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Abstract

The invention provides an enzyme linked immunosorbent assay kit for detecting medroxyprogesterone, which comprises an enzyme label plate coated with a coating antigen, an enzyme marker, a medroxyprogesterone standard substance solution, a substrate coloured solution, a stopping solution, a washing liquid and a reconstitution fluid, the coating antigen is the medroxyprogesterone coupling antigen, and the enzyme marker is an enzymic-labelled antibody. The invention also discloses a method for detecting medroxyprogesterone by using the enzyme linked immunosorbent assay kit, which comprises the following steps: pre-treating a sample, then detecting by the kit, and finally analyzing the detection result. The enzyme linked immunosorbent assay kit can be used for detecting the medroxyprogesterone content in chicken, fish and shrimp samples, operation is simple, cost is low, sensitivity is high, and the kit enables on-site monitor and is suitable for screening a lot of samples.

Description

technical field [0001] The present invention relates to enzyme-linked immunoassay detection technology, in particular to an enzyme-linked immunoimmunoassay kit for detecting medroxyprogesterone, which can qualitatively and quantitatively detect medroxyprogesterone drug in animal tissue (chicken, fish, shrimp, etc.) samples residue. Background technique [0002] Medroxyprogesterone (Medroxyprogesterone), Chinese alias: medroxyprogesterone, medroxyprogesterone, is a synthetic progesterone derivative, and Medroxyprogesterone is a synthetic progestin drug. The drug is a progestogen with strong action, no estrogen activity, and is effective both oral and injection. Its progesterone activity is 20 to 30 times that of progesterone when injected subcutaneously, and 10 to 15 times that of progesterone when taken orally. After intramuscular injection, it is locally stored in the tissue and released slowly, producing a long-acting effect, which can last for more than 2 to 4 weeks, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/64G01N33/577G01N33/543
CPCG01N33/543G01N33/566G01N33/64G01N33/74
Inventor 何方洋扶胜崔海峰冯静聂雯莹陶光灿贾芳芳徐念琴
Owner BEIJING KWINBON BIOTECH
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