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Method measuring N-acetyl-beta-D-amidoglucosaccharase and liquid type stable reagent

A technology of glucosamine and acetylamino, which is applied in the field of N-acetyl-β-D-glucosaminidase determination method and liquid type stabilizing reagent, can solve the problems of inconvenient use time, high use cost, short stability period and the like

Inactive Publication Date: 2005-10-26
商纯尔
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the CPNP continuous monitoring method dry powder assay reagent is reconstituted, the stability period is short (storage at 4°C can only be stable for 7 days), and the user cost is high
PNP fixed-time two-point method assay kit (model: dry powder + buffer solution) has three times the assay sensitivity of CPNP continuous monitoring method, but because the dry powder substrate of the kit is very difficult to dissolve, the actual clinical use time is very inconvenient, and the dry powder The substrate is only stable for about 15 days after reconstitution with buffer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Fixed time two-point method determination reagent, p-nitrobenzene-β-D-N-acetylglucosamine is selected as the substrate for NAG action, the surfactant is preferably F88, and the stabilizer is preferably EDTA-2Na.

[0032] A component:

[0033] P-Nitrobenzene-β-D-N-acetylglucosamine 20mmol / L

[0034] F88 (surfactant) 5g / L

[0035] EDTA-2Na (stabilizer) 50mmol / L

[0036] NaN3 (preservative) 1g / L

[0037] Corbic acid oxidase (interfering agent) 8mmol / L

[0038] Component B:

[0039] p H 4.6 Citric acid-trisodium citrate buffer 50mmol / L

[0040] NaN3 (preservative) 1g / L

[0041] F88 (surfactant) 5g / L

[0042] C component:

[0043] NaOH (stop solution) 100mmol / L

[0044] Before the reagent of Example 1 is used for the measurement, the A component and the B component are mixed in a ratio of 5:1 to form the measurement reagent R1, and the C component is the measurement reagent R2. When measuring the sample, the fixed-time two-point method is used, the temperature is 37℃,...

Embodiment 2

[0045] Example 2: Fixed time two-point method determination reagent, select m-methoxynitrovinyl-β-DN-acetylglucosamine as the substrate for NAG action, the surfactant is preferably emulsifier OP, and the stabilizer is preferably mannose alcohol.

[0046] A component:

[0047] M-Methoxynitrobenzene-β-D-N-acetylglucosamine 16mmol / L

[0048] Emulsifier OP 10ml / L

[0049] Mannitol 12g / L

[0050] PC800 1g / L

[0051]Ascorbic acid oxidase 8mol / L

[0052] Component B:

[0053] p H 4.4 Glycine buffer 80mmol / L

[0054] PC800 1g / L

[0055] Emulsifier OP 10ml / L

[0056] C component:

[0057] P H 10.0 Borax buffer 100mmol / L

[0058] The measurement method of Example 2 is the same as that of Example 1, but the measurement wavelength is adjusted to 505 nm in order to effectively improve the sensitivity and anti-interference ability of the measurement reagent.

Embodiment 3

[0059] Example 3: Continuous monitoring method determination reagent, 6-methyl-2-pyridine-N-acetyl-1-thio-β-D-aminoglucoside is selected as the substrate for NAC action, and the surfactant is preferably Brij35, which is stable The agent is preferably 18-crown 5 ether.

[0060] A component:

[0061] P H 4.4 Acetic acid-sodium acetate buffer 200mmol / L

[0062] PC800 1g / L

[0063] Brij35 8g / L

[0064] Potassium ferrocyanide 1mmol / L

[0065] Grade B:

[0066] 6-Methyl-2-pyridine-N-acetyl-1-thio-β-D-aminoglucoside 12mmol / L

[0067] Brij35 8g / L

[0068] 18-crown 5 ether 200mmol / L

[0069] PC800 1g / L

[0070] When the reagent of Example 3 is used for the measurement, the reagent does not need to be prepared and used directly. The A component is the measurement reagent R1, and the B component is the measurement reagent R2. When measuring the sample, the continuous monitoring method is used, the temperature is 37℃, R1:sample:R2 is 300:20:100, and the measurement wavelength is 340nm. After ...

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PUM

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Abstract

This invention relates to a method of detecting N-acetyl-beta-D-aminoglucosaccharase, and its stable solution. It characters: use its stable solution to automatically carry the detection of N-acetyl-beta-D-aminoglucosaccharase in human fluid by the fixing time two point method and detecting method continuously.

Description

Technical field [0001] The invention relates to a method for determining N-acetyl-β-D-glucosaminidase and the field of liquid-type stabilizing reagents. Background technique [0002] At present, the measurement methods of N-acetyl-β-D-glucosaminidase in clinical applications mainly include: radioimmunoassay, fluorescence analysis, end-point colorimetry, etc. The common characteristics of radioimmunoassay, fluorescence analysis and end-point colorimetry are the need for special equipment, cumbersome operation, long test time, and it is not suitable for clinical large-flow automated analysis. At present, the most widely used domestic determination of urine N-acetyl-β-D-glucosamine (NAG) activity is the CPNA continuous monitoring method determination kit (dry powder) and the PNP fixed time two-point method determination kit (dry powder + buffer). After the CPNP continuous monitoring method dry powder determination reagent is reconstituted, the stability period is short (stable at 4°...

Claims

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Application Information

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IPC IPC(8): C12Q1/34
Inventor 商纯尔
Owner 商纯尔
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