In vitro diagnosis kit of hepatitis B virus E antibody through dual-antigen sandwich method and preparation method thereof

A double-antigen sandwich and in vitro diagnostic technology, which is applied in the field of immunomedicine analysis, can solve the problems of HBeAg unable to be coupled with enzymes, epitope inactivation, HBeAg inactivation, etc., to achieve strong clinical applicability, accurate interpretation of results, and results accurate effect

Inactive Publication Date: 2009-12-09
BEIJING BOSHENGFU BIOTECH CO LTD
View PDF0 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fourth, the color difference between negative and positive is obvious, and it is almost possible to judge the negative and positive results by visual inspection.
When the sodium periodate method is used to couple HBeAg to the enzyme, the amino group within the range of the determinant of the enzyme and HBeAg is more likely to bind to the enzyme
When the enzyme molecule is combined with HBeAg, on the one hand, when the enzyme binds to the amino group on the HBeAg epitope, the epitope will be inactivated; on the other hand, if HBeAg is coupled to more than one enzyme molecule, due to the enzyme The molecular weight is 43,000 Daltons, which is much larger

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • In vitro diagnosis kit of hepatitis B virus E antibody through dual-antigen sandwich method and preparation method thereof
  • In vitro diagnosis kit of hepatitis B virus E antibody through dual-antigen sandwich method and preparation method thereof
  • In vitro diagnosis kit of hepatitis B virus E antibody through dual-antigen sandwich method and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1 prepares double antigen sandwich method hepatitis B virus E antibody in vitro diagnostic kit (ELISA method) of the present invention

[0065] 1. Preparation of microplates

[0066] (1) Coated

[0067] 0.05M carbonate buffer solution with a pH of 9.6 is the coating diluent, which is mixed evenly with HBeAg and then loaded on the microwell plate.

[0068] Specifically, the coating method includes:

[0069] Na 2 CO 3 1.6g

[0070] NaHCO 3 2.9g

[0071] Distilled water 1000ml

[0072] After the buffer solution is completely dissolved and mixed evenly, add 2 mg of HBeAg and mix evenly, and then add 0.1 ml of HBeAg dilution solution to each well of the microplate. Place at 37°C for 12 hours.

[0073] (2) washing plate

[0074] Use saline as the washing solution. After shaking off the HBeAg dilution in the microwell plate, it was washed twice with physiological saline. Finally drain the microplate.

[0075] (3) closed

[0076] 0.01 M pH 7.2 p...

Embodiment 2～4

[0142] Embodiment 2~4 prepare double antigen sandwich method hepatitis B virus E antibody in vitro diagnostic kit (ELISA method) of the present invention

[0143] Except that plastic beads, plastic tubes or magnetic particles were used as carriers, the other methods were the same as in Example 1 to prepare the double-antigen sandwich hepatitis B virus E antibody in vitro diagnostic kit.

Embodiment 5

[0144] Embodiment 5 prepares double-antigen sandwich method hepatitis B virus E antibody in vitro diagnostic kit (ELISA method) of the present invention

[0145] Except that alkaline phosphatase was used as the labeling enzyme of HBeAg, the others were prepared in the same manner as in Example 1 to prepare the double-antigen sandwich hepatitis B virus E antibody in vitro diagnostic kit.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides an in vitro diagnosis kit (an ELISA method) of a hepatitis B virus E antibody through a dual-antigen sandwich method, which comprises the following components: 1) a recombinant HBeAg coated in a solid phase carrier; 2) an enzyme-labeled recombinant HBeAg; and 3) a substrate solution, for example, a chromogenic substrate solution applied to the ELISA, or an enzymatic chemiluminescent substrate solution. Besides, the invention provides a method for preparing the kit, which comprises the following steps: 1) coating the recombinant HBeAg into the solid phase carrier; 2) performing enzyme labeling on the recombinant HBeAg; 3) preparing the substrate solution; and 4) assembling the components into a finished kit. The kit has the advantages of high sensitivity, good specificity, accurate result and strong clinical applicability, and can accurately reflect the amount of the E antibody in the body of a hepatitis B patient.

Description

technical field [0001] The invention relates to the field of immunomedicine analysis, in particular, the invention relates to a hepatitis B virus E antibody in vitro diagnostic kit and a preparation method. Background technique [0002] Hepatitis B Virus belongs to the family of hepatotropic viruses, which can cause acute hepatitis, chronic hepatitis and liver cirrhosis, and is closely related to the occurrence of liver cancer. It is one of the most widespread and deadly viruses in the world today. According to statistics, more than 3.5 billion people in the world are carriers or patients of hepatitis B [Zucherman J.N., et al., J. Infect, 2000, 41(2): 130-136]. China is a high-incidence and endemic area of ​​hepatitis B, with about 120 million hepatitis B patients [Edited by Zhang Jianying, Research Progress in Viral Liver Diseases, First Edition, China Science and Technology Press, 1992]. Moreover, about 1 million people worldwide die each year due to HBV infection [Parki...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N33/576G01N33/545G01N33/52G01N33/535G01N21/31G01N21/76
Inventor 钟士博冯挺财张娜
Owner BEIJING BOSHENGFU BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap