Method for detecting N-acetyl-beta-amino glucosaccharase activity and diagnosis kit therefor

A technology of glucosamine and diagnostic kits, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, color/spectral characteristic measurement, etc. The stability of reagents is not good enough to achieve the effect of low test cost, fast measurement speed, and easy popularization and application

Inactive Publication Date: 2007-07-11
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Description
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  • Application Information

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Problems solved by technology

The former is unsuitable for routine testing in general hospitals because of the expensive equipment required and the high requirements for the preparation of the substrate solution; the latter has been reported. Although the selection of various experimental conditions has its own advantages, some The operation is cumbersome and time-consuming, and some selected conditions or reagents are not stable enough to meet the requirements of clinical testing

Method used

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  • Method for detecting N-acetyl-beta-amino glucosaccharase activity and diagnosis kit therefor
  • Method for detecting N-acetyl-beta-amino glucosaccharase activity and diagnosis kit therefor
  • Method for detecting N-acetyl-beta-amino glucosaccharase activity and diagnosis kit therefor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment one (single agent)

[0035] Prepare the N-acetyl-β-glucosaminidase diagnostic kit according to the following ingredients and dosage:

[0036] Imidazole / hydrochloric acid buffer 50mmol / L,

[0037] Ammonium sulfate 80g / ml,

[0038] 3,4 bisnitrophenyl-β-acetylglucosamine 2mmol / L.

[0039] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0040] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 405nm, test subwavelength 505nm, tested N-acetyl-β-glucosaminidase samples and reagents The volume ratio is 1:25, the reaction direction is forward reaction (rising reaction), the delay time is about 1 minute, the detection time is about 2 minutes, and the theoretical K value is 1409.

[0041] After adding the samples and reagents, let them mix and reac...

Embodiment 2

[0042] Embodiment two (two doses)

[0043] Prepare the N-acetyl-β-glucosaminidase diagnostic kit according to the following ingredients and dosage:

[0044] Reagent I——

[0045] Disodium hydrogen phosphate-citric acid buffer 50mmol / L,

[0046]Glycerin 2mol / L;

[0047] Reagent II——

[0048] Disodium hydrogen phosphate-citric acid buffer 50mmol / L,

[0049] Propylene Glycol 50% v / v,

[0050] 2-Chloro-4-nitrophenyl-β-acetylglucosamine 2mmol / L.

[0051] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0052] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 405nm, test subwavelength 505nm, tested N-acetyl-β-glucosaminidase samples and reagents I. The volume ratio of reagent II is 2:20:5, the reaction direction is forward reaction (rising reaction), the delay time is about 1 minute, the detection time...

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Abstract

The active feature inspection for acetylglucosamidase and its diagnostic agent box uses its fostering of continuous monitoring method, enzymolysising dual chlorineor chlorine acetylglucosamidase reaction, through the absorbance elevating speed of the 405nm, inspecting its active feature. The agent box comprises the bumper solution, synthesized nitrobenzene betaacetylglucosamidase and stabilizer to make dual agent by the diagnostic agent box to reduce cross impact of all kinds of components. Through visible light analyzer, the result is precise, free from outside contamination, easy for promotion.

Description

technical field [0001] The invention relates to the detection of N-acetyl-β-glucosaminidase activity in the field of medicine, in particular to a method for measuring the activity of N-acetyl-β-glucosaminidase by using an enzyme colorimetric method, and the N-acetyl-β-glucosaminidase obtained thereby. -Acetyl-β-glucosaminidase diagnostic kit, which belongs to the technical field of N-acetyl-β-glucosaminidase activity analysis and detection. Background technique [0002] N-acetyl-β-D-glucosaminidase (NAG)) is an acid hydrolase in intracellular lysosomes. Studies have shown that the activity of NAG in body fluids (blood, urine) can be used as an effective auxiliary diagnostic index for various diseases, and the detection of NAG activity in body fluids has important clinical value. NAG can be used as an extremely sensitive indicator of kidney damage. The urine content of patients with acute renal failure caused by shock is extremely high, up to 1200 times that of normal people...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/25G01N21/31C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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