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Hyaluronidase detection method

A technology of hyaluronidase and detection method, applied in the field of hyaluronidase detection, can solve the problems of high selectivity and sensitivity, inability to quantitatively detect hyaluronidase, expensive antibodies and the like

Active Publication Date: 2016-06-15
山西迪安医学检验中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the methods have high selectivity and sensitivity
Turbidity determination requires expensive antibodies and complex washing steps
Viscometry is only suitable for qualitative detection and not sensitive for quantitative detection of hyaluronidase

Method used

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Experimental program
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Effect test

Embodiment 1

[0058] 1) Magnetic Fe 3 o 4 Preparation of nanospheres: 0.5406g of FeCl 3 ·6H 2O (superferric chloride) was dissolved in 20 mL of EG (ethylene glycol), and after magnetic stirring for 30 min, 2.0 g of PVP (polyvinylpyrrolidone, weight average molecular weight 40,000) was added. Then, in a nitrogen atmosphere, the oil bath was heated to 120° C. for 1 h, and then 1.5 g of NaOAc (anhydrous sodium acetate) was added therein, heating was stopped, and stirring was continued for 30 min. The resulting reaction mixture was transferred to a 50 mL reactor at 200° C. for 12 h. After the reaction, cool to 25°C, separate with a magnet, disperse in ethanol and water, and wash to obtain magnetic Fe 3 o 4 nanospheres.

[0059] 2) Magnetic Fe 3 o 4 SiO 2 Preparation of nanospheres: 0.155 g of magnetic Fe 3 o 4 Nanospheres were redispersed in the mixed system of 20mL ethanol and 1mL water, ultrasonically dispersed, then 1mL ammonia water was added, ultrasonicated for 15min, and 5.2ml ...

Embodiment 2

[0065] 1) Magnetic Fe 3 o 4 Preparation of nanospheres: 0.5406g of FeCl 3 ·6H 2 O (superferric chloride) was dissolved in 15 mL of EG (ethylene glycol), and after magnetic stirring for 20 min, 1.5 g of PVP (polyvinylpyrrolidone, weight average molecular weight 3000) was added. Then, in a nitrogen atmosphere, the oil bath was heated to 110° C. for 0.5 h, and then 1 g of NaOAc (anhydrous sodium acetate) was added therein, heating was stopped, and stirring was continued for 0.5 min. The resulting reaction mixture was transferred to a 50 mL reactor at 180° C. for 10 h. After the reaction, cool to 25°C, separate with a magnet, disperse in ethanol and water, and wash to obtain magnetic Fe 3 o 4 nanospheres.

[0066] 2) Magnetic Fe 3 o 4 SiO 2 Preparation of nanospheres: 0.155 g of magnetic Fe 3 o 4 Nanospheres were redispersed in the mixed system of 15mL ethanol and 0.5mL water, ultrasonically dispersed, then added 0.5mL ammonia water, ultrasonicated for 15min, and 5ml of...

Embodiment 3

[0072] 1) Magnetic Fe 3 o 4 Preparation of nanospheres: 0.5406g of FeCl 3 ·6H 2 O (ferric chloride) was dissolved in 25 mL of EG (ethylene glycol), and after magnetic stirring for 40 min, 2.5 g of PVP (polyvinylpyrrolidone, weight average molecular weight 5000) was added. Then, in a nitrogen atmosphere, the oil bath was heated to 130° C. for 1.5 h, and then 2 g of NaOAc (anhydrous sodium acetate) was added therein, heating was stopped, and stirring was continued for 1.5 min. The resulting reaction mixture was transferred to a 50 mL reactor at 220° C. for 14 h. After the reaction, cool to 25°C, separate with a magnet, disperse in ethanol and water, and wash to obtain magnetic Fe 3 o 4 nanospheres.

[0073] 2) Magnetic Fe 3 o 4 SiO 2 Preparation of nanospheres: 0.155 g of magnetic Fe 3 o 4 Nanospheres were redispersed in the mixed system of 25mL ethanol and 1.5mL water, ultrasonically dispersed, then 1.5mL ammonia water was added, ultrasonicated for 15min, and 7ml of ...

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Abstract

The invention discloses a hyaluronidase detection method. The method includes the steps that firstly, FeO4 nanospheres are prepared; secondly, Fe3O4@SiO2 nanospheres are prepared; thirdly, Fe3O4@SiO2@NH2 nanospheres are prepared; fourthly, L-cysteine modified quantum dots of a CdT2@CdS nuclear shell structure are prepared; fifthly, HA-QDs is prepared; sixthly, the Fe3O4@SiO2@NH2 nanospheres are added into HA-QDs to obtain HA-QDs-Fe3O4 composite structures, HAase solutions with the same volume and different concentrations are added into multiple composite structures for fluorescence reaction, then fluorescence intensity is detected, and a work curve is drawn; seventhly, purified normal human body serum is added into the composite structures for fluorescence reaction, then fluorescence intensity is detected, and HAase concentration in serum is calculated according to the work curve. The method has excellent sensitivity on hyaluronidase detection and can avoid background interference.

Description

technical field [0001] The invention relates to a method for detecting hyaluronidase. Background technique [0002] Hyaluronidase, the English name is Hyaluronidase, is an enzyme that can reduce the activity of hyaluronic acid in the body, thereby improving the fluid permeability of tissues. In the body, the production of many malignant tumors is often accompanied by an increase in the content of hyaluronidase, for example, bladder cancer, rectal cancer, etc. Therefore, sensitively detecting the content of hyaluronidase in the human body is of great significance for clinical diagnosis and initial tumor treatment. [0003] Traditional hyaluronidase detection methods include turbidity measurement, viscometry, colorimetry, and the like. However, none of the methods has high selectivity and sensitivity. Turbidity measurements require expensive antibodies as well as complex washing steps. Viscometry is only suitable for qualitative detection, but not sensitive for quantitativ...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/573
CPCG01N21/6486G01N33/573
Inventor 夏云生宫素芹陈露吕扬朱霜霜朱慧刘春秀
Owner 山西迪安医学检验中心有限公司