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Method for detecting glycocholic acid by adopting one-step method

A technology of glycocholic acid and step method, which is applied in the field of medical detection, can solve the problems of long ELISA detection time and unreachable glycocholic acid, and achieves the effects of simple operation and easy reagents.

Pending Publication Date: 2020-03-06
RENMIN UNIVERSITY OF CHINA
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Problems solved by technology

[0003] At present, the main methods for quantitatively detecting the content of glycocholic acid include radioimmunoassay, ELISA and other methods. The radioimmunoassay will release radioactive substances to endanger the environment and operators, while the detection time of ELISA is longer, and the current detection method of glycocholic acid cannot be simple. The "one-step" detection

Method used

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  • Method for detecting glycocholic acid by adopting one-step method
  • Method for detecting glycocholic acid by adopting one-step method
  • Method for detecting glycocholic acid by adopting one-step method

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Embodiment 1

[0034] 1) Modification of the optical waveguide sensing interface

[0035] Immersion of the optical waveguide sensing interface in HF:HNO 3 = 1:3 solution for 30 minutes, remove impurities and connect hydroxyl groups on the surface, clean it with ultrasonic waves and dry it, and connect bifunctional reagents after silanization to modify the coated antigen of glycocholic acid.

[0036] 2) Glycocholic acid antibody-labeled fluorescence

[0037] The glycocholic acid antibody was labeled with Cy5.5 dye, the dialysis bag was placed in a beaker, submerged in ultrapure water, heated in a microwave oven for 10 min, and the hot water was poured out. This operation was repeated three times. Add a certain concentration of antibody into the dialysis bag, and dialyze in 1 L of 0.15M NaCl solution at room temperature for 4 hours. Dialyze against fresh 1 L of 0.15M NaCl solution overnight at 4°C. The next day with 1L 0.1NaHCO 3 The solution was dialyzed for 4h. Cy5.5 was prepared with D...

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Abstract

The invention discloses a method for detecting glycocholic acid by adopting a one-step method. According to the method, the principle of laser-induced fluorescence is utilized, light with a certain wavelength is utilized to excite fluorescent molecules carried by glycocholic acid antibodies specifically combined with the surface of an optical waveguide, the intensity of fluorescent signals is monitored in real time, and real-time detection of glycocholic acid is achieved. Freeze-dried fluorescently-labeled glycocholic acid antibody powder and glycocholic acid react and then are directly detected, and therefore, the method for detecting glycocholic acid by adopting a one-step method has the advantages that reagents are easy to store, operation is easy and convenient, and portability is achieved, and the content of glycocholic acid in serum is detected through adoption of the one-step method.

Description

technical field [0001] The invention belongs to the field of medical detection, and relates to a method for detecting glycocholic acid, in particular to a one-step method for detecting glycocholic acid. Background technique [0002] Glycocholic acid is one of the conjugated cholic acids formed by the combination of cholic acid and glycine. In the medical field, the content of Glycocholic acid can reflect the health status of the liver. Glycocholic acid in serum can reflect the function of liver cells and the circulation function of hepatobiliary substances. lower. When human liver cells are damaged or bile is stagnant, the metabolism will be disturbed, the content of glycocholic acid absorbed by cells will decrease, and the content of glycocholic acid in the blood will increase. The degree of increase is related to the degree of damage to liver cells. [0003] At present, the main methods for quantitatively detecting the content of glycocholic acid include radioimmunoassay...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402
Inventor 龙峰刘佳瑶徐文娟方顺燕宋丹
Owner RENMIN UNIVERSITY OF CHINA
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