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
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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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Application Information
- IPC
- G01N21/25; G01N21/31; C12Q1/26
- Inventors
- 王尔中
