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Method for detecting 19-nortestosterone and special enzyme-linked immune reagent kit thereof

An enzyme-linked immunosorbent reagent and a technology of nortestosterone, which is applied in the field of detection of 19-nortestosterone, can solve the problems of cumbersome process, complicated equipment, unsuitability for on-site monitoring and screening of a large number of samples, and achieve simple sample pretreatment process, The effect of high accuracy and convenient inspection method

Inactive Publication Date: 2008-12-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The chemical methods for detecting the residual amount of 19-nortestosterone mainly include thin-layer chromatography (TLC), gas chromatography (GC), high-performance liquid chromatography (HPLC), gas-mass spectrometry (GC / MS), and liquid-mass chromatography. On-line (HPLC / MS), capillary electrophoresis (CE), etc., are not suitable for on-site monitoring and screening of a large number of samples due to the complexity of the equipment required and the cumbersome process

Method used

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  • Method for detecting 19-nortestosterone and special enzyme-linked immune reagent kit thereof
  • Method for detecting 19-nortestosterone and special enzyme-linked immune reagent kit thereof
  • Method for detecting 19-nortestosterone and special enzyme-linked immune reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of 19-nortestosterone hapten and carrier protein as a coating source

[0035] The ELISA kit using the conjugate of 19-nortestosterone hapten and carrier protein as the coating source includes:

[0036] (1) a microtiter plate coated with 19-nortestosterone and carrier protein conjugate;

[0037] (2) Alkaline phosphatase-labeled goat anti-mouse anti-antibody working solution: Dilute the alkaline phosphatase-labeled goat anti-mouse anti-antibody with a diluent to an enzyme-labeled anti-antibody working solution with a protein concentration of 0.1-1 μg / L Liquid, 12ml / bottle, 1 bottle. The diluent contains 0.1‰glycerol (which can prevent the enzyme markers from freezing at -20°C and maintain the biological activity of the enzyme markers for a long time), 1% calf serum, and 1% thimerosal preservative (easy to store). )The solution.

[0038] (3) 19-nortestosterone stand...

Embodiment 2

[0093] Example 2. ELISA kit using goat anti-rabbit anti-antibody as coating source and its preparation method

[0094] The ELISA kit with goat anti-rabbit anti-antibody as the coating source includes:

[0095] (1) ELISA plate coated with goat anti-rabbit anti-antibody;

[0096] (2) Alkaline phosphatase-labeled 19-nortestosterone hapten working solution: Dilute the alkaline phosphatase-labeled 19-nortestosterone hapten to a working solution of 0.1-1 μg / L, 12ml / bottle, 1 bottle. The diluent used is a solution containing 0.1‰ glycerol (which can prevent the enzyme markers placed in the -20°C environment from freezing, and can also maintain the biological activity of the enzyme markers for a long time), and 1% thimerosal preservative (easy to preserve).

[0097] (3) 19-nortestosterone standard solution: 6 bottles of 19-nortestosterone series standard solution, 0μg / L, 0.1μg / L, 0.3μg / L, 0.9μg / L, 2.7μg / L, 8.1μg / L L, 3ml / bottle. The diluent for diluting 19-nortestosterone into a s...

Embodiment 3

[0123] Embodiment 3, test kit precision, accuracy and shelf life test

[0124] 1. Kit precision test

[0125] (1) Precision test of standard product

[0126] Three batches of the kits prepared in Example 1 and Example 2 were taken respectively for the precision experiment, and 10 kits were taken from each batch of kits, and 20 microwells were respectively extracted from the enzyme-linked plate of each kit. Measure the absorbance value (OD value) of the 0.9 μg / L standard solution, and calculate the coefficient of variation. The measurement results of the three batches of kits in Example 1 are shown in Table 1, and the results show that the coefficient of variation ranges from 5.4% to 10.9%. Meets the requirement for a precision of less than 15%.

[0127] Table 1 Standard repeatability test

[0128]

[0129] The measurement results of the three batches of kits in Example 2 are shown in Table 2, and the results show that the coefficient of variation ranges from 5.9% to 10.6%...

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Abstract

The present invention discloses a method for detecting 19-nandrolone and its special-purpose ELIA kit. Said kit includes 19-nandrolone specific antibody, coating source and enzyme label. The described coating source is conjugate of 19-nandrolone semiantigen and carrier protein or antiantibody. The described enzyme label is enzyme-labelled antiantibody or enzyme-labelled 19-nandrolone semiantigen. When the described coating source in the conjugate of 19-nandrolone semiantigen and carrier protein, the described enzyme label is enzyme-labelled antiantibody, and when the described coating source is anti-antibody, the described enzyme label is the enzyme-labelled 19-nandrolone semi-antigen.

Description

technical field [0001] The invention relates to a method for detecting 19-nortestosterone and a special ELISA kit. Background technique [0002] 19-nortestosterone (19-nortestosterone, NT) is a synthetic hormone that has androgenic effects and belongs to anabolic steroids. Clinically, it is mainly used to treat anemia and senile osteoporosis. It can also promote muscle growth and development, increase training endurance and training load. However, it also has very serious side effects. Ingestion by the mother will masculinize the female fetus and cause reproductive deformities. Long-term use by women will produce masculine features, and long-term use by men will cause premature baldness. Short-term high-dose use can lead to liver dysfunction, and may also lead to cancer, diabetes, mental disorders, etc. Therefore, it is necessary to strengthen the detection of this drug. [0003] The chemical methods for detecting the residual amount of 19-nortestosterone mainly include ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠何方洋万宇平张素霞冯才伟史为民吴小平汪善良江海洋
Owner CHINA AGRI UNIV
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