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Method for rapidly detecting urine steroid hormone by adopting immunochemiluminometric assay

A technology of steroid hormone and immunochemistry, which is applied in the field of analytical chemistry and medicine, can solve the problems of single type of steroid hormone, equipment limitation, application limitation, etc., and achieve cheap and easy reagents, low cost, wide detection range and accuracy Effect

Inactive Publication Date: 2016-12-07
WUXI NO 4 PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although urosteroid hormones can reflect the average level in a certain period of time in the body, and urine sample collection has the advantages of non-invasive and convenient, but due to the lack of relevant literature reports, the single type of steroid hormones studied, cumbersome detection or equipment limitations, etc. The application is limited in the diagnosis and treatment of diseases such as clinical dysplasia

Method used

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  • Method for rapidly detecting urine steroid hormone by adopting immunochemiluminometric assay
  • Method for rapidly detecting urine steroid hormone by adopting immunochemiluminometric assay
  • Method for rapidly detecting urine steroid hormone by adopting immunochemiluminometric assay

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Effect test

Embodiment 1

[0020] The invention is used to detect five kinds of steroid hormones in women's urine.

[0021] Take 500 μL of fresh overnight morning urine from women, centrifuge at 10,000 rpm / min for 5 minutes, and store the supernatant at -20°C. Accurately measure 0.5 mL of urine sample into a 5 mL glass graduated tube, add 1.0 mL of acetic acid-sodium acetate buffer solution (pH 5.0) and 1800-2200 U of β-glucuronidase aqueous solution, mix well, and place at 50-60 °C Enzymolysis in a water bath for 3h. After the graduated tube was cooled to room temperature, it was diluted to within the detection range of the instrument for ICMA detection. The quality control was calibrated with E3, HC, 17β-E2, T and P before the determination of urine samples, and the data was processed using the software that comes with the ICMA instrument. The standard solution matrix S0 was used as the zero standard point.

Embodiment 2

[0023] The invention is used to detect five kinds of steroid hormones in male urine.

[0024] Take 500 μL of fresh overnight morning urine from men, centrifuge at 10,000 rpm / min for 5 min, and store the supernatant at -20°C. Accurately measure 0.5 mL of urine sample into a 5 mL glass graduated tube, add 1.0 mL of acetic acid-sodium acetate buffer solution (pH 5.0) and 1800-2200 U of β-glucuronidase aqueous solution, mix well, and place at 50-60 °C Enzymolysis in a water bath for 3h. After the graduated tube was cooled to room temperature, it was diluted to within the detection range of the instrument for ICMA detection. The quality control was calibrated with E3, HC, 17β-E2, T and P before the determination of urine samples, and the data was processed using the software that comes with the ICMA instrument. The standard solution matrix S0 was used as the zero standard point.

Embodiment 3

[0026] Evaluation of standard addition recovery rate for detection of urinary total steroid hormones by using the present invention.

[0027] Under the experimental conditions determined by this method, after the five steroid hormones are pretreated, the total steroid hormone concentration after enzymatic hydrolysis is detected; different urine samples are added with different concentrations of a certain steroid hormone standard solution to detect a certain concentration The recovery rate of steroid hormones, that is, adding low, medium, and high concentrations of steroid hormone standard solutions to different urine samples, using ICMA to detect the concentration of the standard sample, and comparing it with a known amount of standard substance, repeat The test was performed 6 times, and the calculated average recovery rate of the urine sample is shown in Table 2. E. 3 , HC, 17β-E 2 , T and P average recoveries range between 93.7% to 117.9%, indicating that the method of th...

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Abstract

The invention relates to a method for rapidly detecting a urine steroid hormone by adopting an immunochemiluminometric assay and belongs to the fields of analytical chemistry and medical science. According to the method, a beta-glucuronidase enzymolysis method is combined with the immunochemiluminometric assay to rapidly detect concentrations of five steroid hormones, namely urinary estriol, hydrocortisone, 17beta-estradiol, testosterone and progesterone, at the same time, and limit of quantitation is 0.017-4ng / mL. The method provided by the invention is rapid, sensitive and accurate, the requirement of detecting the five steroid hormones of human urine at the same time can be met, the used reagent is cheap and available, cost is low, and research and development of sample enzymolysis and immunochemiluminometric assay integrated equipment can be facilitated, so that the method provided by the invention has a broad application prospect.

Description

technical field [0001] The invention relates to a method for rapidly detecting urinary steroid hormones by immunochemiluminescence (ICMA), belonging to the fields of analytical chemistry and medicine. Background technique [0002] Steroid hormones mainly include adrenal cortex hormones and sex hormones (including male hormones and anabolic hormones, progesterone and estrogen, etc.), which play an extremely important role in maintaining life, body development, sexual function regulation, immune regulation and birth control. . Over the years, estriol (E 3 ), hydrocortisone (HC), 17β-estradiol (17β-E 2 ), testosterone (T) and progesterone (P) and other blood steroid hormones have become common clinical monitoring indicators for the diagnosis and treatment of many human diseases such as endocrine diseases. It only represents the instantaneous level of steroid hormones in the human body, and the invasiveness of blood sample collection is often one of the main reasons affecting...

Claims

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Application Information

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IPC IPC(8): G01N33/74
CPCG01N33/74
Inventor 徐庄剑马亚萍夏冰洋
Owner WUXI NO 4 PEOPLES HOSPITAL
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