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 t

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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