Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody

An enzyme-linked immunosorbent assay and enterovirus technology, which is used in the detection of enterovirus type 71 antibodies by enzyme-linked immunosorbent assay, and the qualitative or quantitative detection of enterovirus type 71-specific antibodies, which can solve the problem of poor sensitivity and specificity. , the test cycle becomes longer, etc., to reduce the effect of false positives

Inactive Publication Date: 2012-07-04
ZHEJIANG PUKANG BIOTECH
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most commonly used method is still the neutralization experiment, but the sensitivity and specificity of this method are poor
Moreover, this method requires cell culture durin

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
  • Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody
  • Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody
  • Enzyme-linked immunosorbent assay method for enterovirus 71-specific antibody

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Example 1

[0034] Detection of EV71 antibody titer in mouse serum by immobilized and purified EV71 virus antigen

[0035] 1) Dilute the purified EV71 virus antigen to 2 μg / ml with carbonate buffer (pH 9.6), add 100 μl / well to the wells of the microtiter plate, incubate overnight at 4°C, wash with PBS Wash the plate 3 times with liquid;

[0036] 2) Add 200 μl / well of blocking solution to the wells of the enzyme-labeled plate, incubate at 37°C for 1 hour, and wash the plate 3 times with PBS washing solution;

[0037] 3) Add 5% sucrose to the wells of the enzyme-labeled plate at an amount of 100 microliters / well, incubate at 37°C for 0.5 to 1 hour, pat dry, and store at 4°C until use;

[0038] 4) Dilute the test serum 10 times with 0.9% normal saline, and then use PBS containing 1% BSA to make a series of dilutions of the serum: 1:20, 1:40, 1:80, 1:160, 1: 320, 1:640, 1:1280, 1:2560, add 100 microliters / well to the wells of the pre-prepared ELISA plate, incubate at 37°C for 1 hou...

Example Embodiment

[0055] Example 2

[0056] Detection of EV71 antibody positive rate in mouse serum with fixed and purified EV71 virus antigen

[0057] 1) Dilute the purified EV71 virus antigen to 3 μg / ml with carbonate buffer (pH 9.6), add 100 μl / well to the wells of the ELISA plate, incubate overnight at 4°C, wash with PBS Wash the plate 3 times with liquid;

[0058] 2) Add 200 μl / well of blocking solution to the wells of the enzyme-labeled plate, incubate at 37°C for 1 hour, and wash the plate 3 times with PBS washing solution;

[0059] 3) Add 100 μl / well of 5% sucrose to the wells of the enzyme-labeled plate, incubate at 37°C for 0.5 to 1 hour, pat dry, and store at 4°C until use

[0060] 4) Dilute 75 mouse positive sera by 100 times, add 100 microliters / well to the wells of the pre-prepared ELISA plate, incubate at 37°C for 1 hour, wash the plate with PBS washing solution 4 times, and set up 2 negative control wells and 1 blank well;

[0061] 5) Add an enzyme-labeled antibody to each well of the EL...

Example Embodiment

[0070] Example 3

[0071] Detection of EV71 antibody positive rate in mouse serum by fixed virus-specific epitope polypeptide

[0072] 1) Dilute the purified EV71 virus antigen to 3 μg / ml with carbonate buffer (pH 9.6), add 100 μl / well to the wells of the ELISA plate, incubate overnight at 4°C, wash with PBS Wash the plate 3 times with liquid;

[0073] 2) Add 200 μl / well of blocking solution to the wells of the enzyme-labeled plate, incubate at 37°C for 1 hour, and wash the plate 3 times with PBS washing solution;

[0074] 3) Add 100 μl / well of 5% sucrose to the wells of the enzyme-labeled plate, incubate at 37°C for 0.5 to 1 hour, pat dry, and store at 4°C until use

[0075] 4) Dilute 75 parts of mouse serum inoculated with EV71 virus by 50 times, add 100 microliters / well to the wells of the pre-prepared ELISA plate, incubate at 37°C for 1 hour, and wash the plate with PBS washing solution 4 times , Set up 2 negative control wells and 1 blank well at the same time;

[0076] 5) Add an ...

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 relates to the field of biotechnology, in particular to an enzyme-linked immunosorbent assay method for an enterovirus 71 (EV71 for short)-specific antibody in serum. Through a method of indirect enzyme-linked immunosorbent assay (ELISA), a prepared EV71 viral antigen is immobilized on an ELISA plate, and the antibody is detected through specific adsorption of antigen and antibody. Since the coated antigen comprises a specific antigen epitope for the EV71 virus, the method can be used for detecting specific antibody for the EV71 virus in the serum, and false positive of serum detection is reduced. The enzyme-linked immunosorbent assay method for the enterovirus 71-specific antibody is suitable for detecting a large batch of animal or human serum samples, and is suitable for large-scale serological and epidemiological investigation.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to the qualitative or quantitative detection of enterovirus 71 (Enterovirus 71, referred to as EV71) specific antibody, and the detection method of enterovirus 71 antibody by enzyme-linked immunosorbent assay. Background technique [0002] Infectious diseases caused by enteroviruses mostly occur in children under 5 years old, and can cause herpes on hands, feet, mouth and other parts. The main pathogens causing HFMD are Enterovirus 71 and Coxsackievirus A16. A large number of epidemiological studies have shown that enterovirus 71 infection mostly occurs in infants under the age of 5, and its infection may cause a variety of serious diseases related to the nervous system, including fatal encephalitis, aseptic meningitis and acute paralysis. Therefore, the specific diagnosis of EV71 virus has very important clinical significance. [0003] The EV71 virus is a member of the Enterovirus g...

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): G01N33/68G01N33/543G01N33/535
Inventor 高孟罗永能周康凤姜云水毛子旭王一虎唐彩华朱莲高丽美庄昉成毛子安毛江森
Owner ZHEJIANG PUKANG BIOTECH
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