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Kit and method for clinically detecting serum or plasma free fatty acid

A free fatty acid and kit technology, applied in the field of testing reagents and biochemistry, can solve the problems of narrowing the linear range of the kit, decreased reactivity, narrow linear range, etc., to improve stability and sensitivity, improve stability, linearity and so on. wide range of effects

Active Publication Date: 2013-07-31
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] (3) The linear range is narrow, mainly because the activity of the chromogen used in step (3) used in the prior art is interfered by coenzyme A and its activity decreases
At present, the Trinder reaction chromogens used in the determination of free fatty acids in the market mainly include MEHA, TBHB, TOOS, etc. These chromogens are greatly interfered by coenzyme A, and the amount of coenzyme A in the reaction must be excessive, so these make these The reactivity of the chromogen is reduced, which narrows the linear range of the kit detection

Method used

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  • Kit and method for clinically detecting serum or plasma free fatty acid
  • Kit and method for clinically detecting serum or plasma free fatty acid
  • Kit and method for clinically detecting serum or plasma free fatty acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Preparation of sulfhydryl reagent:

[0064]

[0065] In a clean glass container, first add the HEPES buffer solution (GOOD's buffer solution is zwitterionic buffer solution, including HEPES buffer solution, MES buffer solution, MOPS buffer solution, etc.) that has been weighed according to the formula ratio, and then add 300ml / L in sequence NEM, 600ml / L DTNB, 40ml / L EDTA, 50ml / L MIT, add appropriate amount of water, add 5ml / L Triton X-100, adjust the pH to pH 6~9 with concentrated hydrochloric acid, the dosage is about 5ml / L, and then stir After filtration, distilled water was added to the obtained filtrate to quantify.

[0066] Preparation of Acyl-CoA Synthetase and Acyl-CoA Oxidase:

[0067] (1) Preparation of acyl-CoA synthetase: culture MK11-1 strain, collect cells, sonicate, centrifuge to get supernatant, precipitate with 30-70% ammonium sulfate, isoelectric point precipitation, DEAE-Cellulose32 column chromatography, molecular sieve After chromatography, it w...

Embodiment 2

[0076] Preparation of sulfhydryl reagent:

[0077]

[0078] Preparation method is with embodiment 1.

[0079] The preparation of acyl-CoA synthetase and acyl-CoA oxidase is the same as in Example 1.

[0080] A kit for clinical detection of free fatty acids in serum or plasma, consisting of the following two reagents:

[0081] Reagent 1:

[0082]

[0083]

[0084] Reagent 2:

[0085]

Embodiment 3

[0087] Preparation of sulfhydryl reagent:

[0088]

[0089] Preparation method is with embodiment 1.

[0090] The preparation of acyl-CoA synthetase and acyl-CoA oxidase is the same as in Example 1.

[0091] A kit for clinical detection of free fatty acids in serum or plasma, consisting of the following two reagents:

[0092] Reagent 1:

[0093]

[0094] Reagent 2:

[0095]

[0096] When measuring the sample, use the two-point endpoint method, the temperature is 37°C, the dominant wavelength is 550nm, the reaction direction is upward (positive reaction), and the sample addition steps are as follows:

[0097] Sample 4μL

[0098] Reagent 1 200μL

[0099] Mix well, incubate at 37°C for 3-5min, and read the absorbance value A1

[0100] Reagent 2 50μL

[0101] Mix well, incubate for 3-5min, read absorbance A2

[0102] ΔA 样本 =A2-A1

[0103]

[0104]

[0105] Using a fully automatic biochemical analyzer, the instrument will automatically give the concentration ...

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Abstract

The invention discloses a kit and method for clinically detecting serum or plasma free fatty acid. The kit is composed of a reagent 1 and a reagent 2, wherein the reagent 1 comprises a buffer, ATP, a coenzyme A, 4-amino antipyrine, an ion activator, an acyl coenzyme A synthetase, stabilizer, a preservative, a surfactant, and concentrated hydrochloric acid, and the reagent 2 comprises a buffer, a chromogen, a composite sulfhydryl reagent, a peroxidase, an acyl coenzyme A oxidase, a stabilizer, a preservative, a surfactant and concentrated hydrochloric acid. The kit has the advantages of good stability, high specificity of the free fatty acid, and wide linear range. The serum or plasma free fatty acid is detected by the enzyme method, and the method has the advantages of high precision, andsmall measuring error.

Description

technical field [0001] The invention relates to the technical fields of biochemistry and test reagents, in particular to a kit for clinically detecting serum or plasma free fatty acids (NEFA) and a method thereof. Background technique [0002] Free fatty acids (FFA), also known as non-estered fatty acids (NEFA), are the hydrolysis products of fat in the human body. Under the action of lipase, triglycerides release free fatty acids, which circulate in the blood and are combined with albumin for transport. Free fatty acids serve as sources of metabolic energy, substrates for cell membrane structure, and precursors for many intracellular signaling molecules (such as prostaglandins) in humans. Although they only account for a small part of body fat, they are one of the important energy substances of the human body, meeting a large part of the energy demand, and are closely related to glucose metabolism and fat metabolism. [0003] Abnormal metabolism of free fatty acids plays ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/28C12Q1/26C12Q1/25C12R1/38
Inventor 邹炳德贾江花
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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