Human serum glycated albumin array kit

A technology of glycosylated albumin and a detection kit, which is applied in the field of medical testing, can solve problems such as reaction interference, poor stability of the kit, and poor stability, so as to reduce the amount of protease and ascorbate oxidase, reduce the cost of enzyme production, and improve stability Effect

Active Publication Date: 2012-07-11
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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AI Technical Summary

Problems solved by technology

[0007] (1) The interference of globulin cannot be completely ruled out: the protease substrate used has poor specificity and can decompose all protein types in serum, so the interference of globulin cannot be ruled out;
[0008] (2) The stability of the kit is poor: because the coupling is a Trinder reaction, and this reaction is easily interfered by reducing substances. Due to the current large input of reducing substances such as ascorbic acid, the high reducing substances in the serum of many patients have a great impact on the reaction. interference; and the ascorbic acid oxidase used to remove ascorbic acid is easily decomposed by protease; protease itself can also undergo self-decomposition reaction, and the stability is poor;
[0009] (3) The cost of the kit is high: in order to completely decompose glycated albumin into glycated amino acids so as to participate in the next step reaction, a high concentration of protease is used, and the amount of enzyme used is also high, so the overall cost of the kit is very high;
[0010] (4) The existing glycated albumin on the market does not give a reference range, only the reference range of glycated albumin / albumin is given, which has certain defects for clinical guidance

Method used

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  • Human serum glycated albumin array kit
  • Human serum glycated albumin array kit
  • Human serum glycated albumin array kit

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Embodiment Construction

[0049] The present invention will be further described in detail below in conjunction with the examples, but not limited thereto.

[0050] A human serum glycated albumin detection kit, comprising reagent 1 and reagent 2, wherein:

[0051]Reagent 1:

[0052]

[0053] Reagent 2:

[0054]

[0055] Proteinase K co-expression body preparation:

[0056] ① Whole gene synthesis of protease co-expression body: find the protease K gene sequence of Tritirachium album limber from GENBANK, add 30 bp nucleotides at the 5' end of the gene sequence, including the enzyme cutting site NdeI, The stop codon was removed from the 3' end, and 120 bp nucleotides were added to the 5' end of the cucumber-derived (Cucumis sp.) ascorbate oxidase gene sequence. The stop codon was also removed from the 3' end, and the restriction site XhoI was added. Carry out whole gene synthesis together;

[0057] ②The recombinant gene is connected into the simple vector of cloning vector pMD-18-T;

[0058] ③I...

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Abstract

The invention provides a human serum glycated albumin assay kit which comprises a reagent 1 and a reagent 2, wherein the reagent 1 comprises the following components: 20-200mmol / L of buffer solution, 5-50KU / L of protease co-expression vector, 10-50KU / L of peroxisome, 2-20mmol / L of 4-amino antipyrine, 0.01-1% of preservative and 0.01-1% of stabilizer, and the protease co-expression vector is obtained by cloning a protease gene and an ascorbic acid oxidase gene onto a same vector for performing co-expression; and the reagent 2 comprises the following components: 20-200mmol / L of buffer solution, 5-50U / mL of fructose lysine enzyme, 1-10mmol / L of N, N-bis(4-sulfobutyl ether)-3-methylaniline, 0.01-1% of preservative and 0.01-1% of stabilizer. The kit can remove the interferences of globulin and ascorbic acid in human serum and has good stability and low cost.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a detection kit for human serum glycosylated albumin. Background technique [0002] Diabetes is a combination of sugar, protein, fat, water, and A series of metabolic disorder syndromes such as electrolytes are one of the major diseases that endanger human diseases. [0003] Albumin in human serum accounts for 66% of serum protein, and the rest is immunoglobulin; non-enzymatic glycosylation reaction occurs between glucose in serum and the N-terminus of serum protein to generate glycosylated serum protein, and glycosylated albumin accounts for glycosylation More than 90% of serum protein. In addition, since the content of albumin exists stably in serum, while globulin will fluctuate due to infection and other disease factors; therefore, accurate determination of glycosylated albumin is very important to accurately reflect the blood sugar control of diabetes. [0004] Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 邹炳德邹继华贾江花沃燕波徐秀芬
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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