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Direct quantitative detection kit for alpha-ketoglutaric acid in human serum

A technology for quantitative detection of ketoglutaric acid, applied in the detection field, can solve problems such as high cost, cumbersome detection method, and inability to directly measure, and achieves low cost, improved operation repeatability, and guaranteed operability and safety. Effect

Inactive Publication Date: 2017-05-24
EAST CHINA UNIV OF SCI & TECH
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  • Abstract
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Problems solved by technology

[0006] The purpose of the present invention is to provide a direct quantitative detection kit for α-ketoglutarate in human serum, which aims to overcome the deficiencies in the prior art such as inability to directly measure, cumbersome detection methods, high cost, etc., thereby greatly expanding its Application in clinical diagnosis

Method used

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  • Direct quantitative detection kit for alpha-ketoglutaric acid in human serum

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Experimental program
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Effect test

Embodiment 1

[0030] 1. Collection and processing of serum samples

[0031] In patients with liver cirrhosis and liver cancer, 5 mL of venous blood was drawn in the morning without anticoagulation. It was stored in a refrigerator at 0°C for 60 minutes. After taking it out, it was centrifuged at 5000 rpm for 15 minutes.

[0032] 2. Detection of α-ketoglutarate in human serum by α-ketoglutarate kit

[0033] a) Parameter setting. Put the black 96-well plate into the carrier, directly set Read→Test method Fluorescence intensity→Detection method endpoint / kinetic detection→Optical device type Filters→Ok→A new parameter will appear, that is, according to α-ketoglutarate detection The nature of the needle selects an excitation wavelength of 530nm, an emission wavelength of 590nm and a gain value of 50. Start testing after setting.

[0034] b) Detect the concentration of α-ketoglutarate in serum. After thawing the serum sample and the kit solution in turn, pour them into the holes and place them on the l...

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Abstract

The invention relates to the field of detection technology and particularly discloses a direct quantitative detection kit for alpha-ketoglutaric acid in human serum. The direct quantitative detection kit comprises a serum sub-packaging tube, a hepes buffer solution, a reducing coenzyme, an enzyme stabilizer, a fluorescent dye and an alpha-KG standard. The kit disclosed by the invention can quickly and efficiently detect alpha-ketoglutaric acid in the serum of patients suffering from liver cirrhosis and liver cancer, thereby providing a technical guarantee for detecting and screening patients suffering from early liver cirrhosis and liver cancer. Compared with the traditional chromatography coupled detection method, nuclear magnetic resonance method and the like, the kit disclosed by the invention is low in cost and simple to operate and has the characteristics of high speed and high throughput; the kit for detecting alpha-ketoglutaric acid in serum has specific selectivity and relatively high stability; and compared with a biological detection method, the kit is obviously improved in operation repeatability, can be used as an effective tool for diagnosing liver cirrhosis and liver cancer, is of great significance to early detection of liver diseases and evaluation of treatment effects, and has broad application prospects.

Description

Technical field [0001] The invention relates to the technical field of detection, in particular to a direct quantitative detection kit for the important small molecule biomarker α-ketoglutarate in human serum. Background technique [0002] α-Ketoglutaric acid (α-Ketoglutaric acid, abbreviated as α-KA, CAS: 328-50-7) is an important biological compound, and its usual form is its anion, namely α-ketoglutaric acid Salt (α-ketoglutarate, referred to as α-KG). Alpha-ketoglutarate is a key substance in the tricarboxylic acid cycle. It is located after isocitrate and before succinyl-CoA in the cycle. The amino groups of amino acids are bound to α-ketoglutarate through transamination, and as the blood is carried to the liver, ammonia enters the urea cycle in the liver, so α-ketoglutarate is widely present in human serum. Existing research results have shown that changes in the concentration of α-ketoglutarate in the blood are closely related to many diseases, such as non-alcoholic fatt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/574
CPCG01N33/582G01N33/57438G01N33/57488G01N2800/085
Inventor 朱为宏何业郭志前沈超宋熊荣陈帆唐俊马石丽敏张鹏马亚光田禾
Owner EAST CHINA UNIV OF SCI & TECH
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