Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Kit for detecting liver cancer, hepatitis and/or liver cirrhosis and application of kit in AKR1B10 and AFP combined quantitative determination

A 1B10, kit technology, applied in the field of medicine and biology, can solve problems such as low concentration, achieve the effects of strong fluorescence signal, improve sensitivity and specificity, and reduce detection time and cost

Pending Publication Date: 2020-06-05
湖南莱拓福生物科技有限公司
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, not all liver cells of patients with liver cancer secrete AFP, and about 30% to 40% of patients still have negative or low levels of AFP, which has obvious limitations.

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
  • Kit for detecting liver cancer, hepatitis and/or liver cirrhosis and application of kit in AKR1B10 and AFP combined quantitative determination
  • Kit for detecting liver cancer, hepatitis and/or liver cirrhosis and application of kit in AKR1B10 and AFP combined quantitative determination
  • Kit for detecting liver cancer, hepatitis and/or liver cirrhosis and application of kit in AKR1B10 and AFP combined quantitative determination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Preparation of AKR1B10 and AFP assay kits.

[0031] 1. The preparation method of the coated microtiter plate is as follows:

[0032]Use 10mmol / L Tris•HCl (pH7.5±0.1) coating buffer to prepare a mixed solution of 5ug / mL rabbit anti-AKR1B10 polyclonal antibody and 10ug / mL mouse anti-AFP monoclonal antibody, and inject the enzyme label at 200uL per well Place in the microwells of the plate for coating reaction at 2-8°C for 12-15 hours, then discard the coating mixed solution. Use 10mmol / L Tris•HCl (pH7.5±0.1), 1% BSA blocking buffer, inject 400uL per well into the microwells of the microplate, place at 37°C for blocking reaction for 2 hours, then discard the blocking buffer liquid, dry naturally.

[0033] 2. The preparation method of the calibrator mixed solution is as follows:

[0034] It is prepared by dissolving AKR1B10 antigen and AFP antigen in sample buffer at a certain concentration. The working concentrations of AKR1B10 are 0, 375, 750, 1500, 3000, 6...

Embodiment 2

[0048] Example 2. Establishing a calibration curve with AKR1B10 and AFP assay kits

[0049] The steps to establish the calibration curve are as follows:

[0050] (1) Add 100ul calibrator to the microwells of the ELISA plate, incubate with shaking at room temperature for 1 hour, add 300ul cleaning solution to each well, and wash repeatedly 3 times;

[0051] (2) Add 100ul AKR1B10 detection antibody and 100ul AFP detection antibody to the microwells of the microplate at the same time, incubate with shaking at room temperature for 1 hour, add 300ul cleaning solution to each well, and wash repeatedly 3 times;

[0052] (3) Add 200ul enhancement solution to each well, shake and incubate at room temperature for 5 minutes;

[0053] (4) Use a time-resolved immunofluorescence analyzer to detect the fluorescence value of europium ions at a wavelength of 340nm and the fluorescence value of terbium ions at a wavelength of 295nm, respectively;

[0054] (5) Data analysis:

[0055] A standa...

Embodiment 3

[0056] Example 3. Using AKR1B10 and AFP assay kits to detect the contents of AKR1B10 and AFP proteins in serum samples of normal people and patients with liver cancer

[0057] (1) Add 100ul serum samples to be tested in the microwells of the ELISA plate, incubate with shaking at room temperature for 1 hour, add 300ul cleaning solution to each well, and wash repeatedly 3 times;

[0058] (2) Add 100ul AKR1B10 detection antibody and 100ul AFP detection antibody to the microwells of the microplate at the same time, incubate with shaking at room temperature for 1 hour, add 300ul cleaning solution to each well, and wash repeatedly 3 times;

[0059] (3) Add 200ul enhancement solution to each well, shake and incubate at room temperature for 5 minutes;

[0060] (4) Use a time-resolved immunofluorescence analyzer to detect the fluorescence value of europium ions at a wavelength of 340nm and the fluorescence value of terbium ions at a wavelength of 295nm, respectively;

[0061] (5) Data...

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 belongs to the technical field of medical biology, and discloses a determination kit for simultaneously detecting contents of aldehyde ketoreductase 1B10 and alpha fetoprotein by using atime-resolved fluorescence immunoassay method, which can be used for screening, diagnosing, curative effect judging, prognosis evaluating or recurrence monitoring of diseases such as liver cancer, hepatitis, liver cirrhosis and the like. The kit comprises an elisa plate, a calibrator, a europium-labeled AKR1B10 detection antibody, a terbium-labeled AFP detection antibody and an enhancement solution. The AKR1B10 and the AFP in the serum to be detected are respectively combined with the AKR1B10 and the AFP coated antibody of the elisa plate, then two detection antibodies are added to form a double-antibody sandwich compound, an enhancement solution is added to dissociate europium ions and terbium ions on the compound, and high-intensity fluorescence is respectively emitted at wavelengths of340nm and 295nm. The fluorescence intensity is in direct proportion to the concentrations of AKR1B10 and AFP in the sample. By adopting the kit, the contents of the two markers can be detected at thesame time, and the detection sensitivity and specificity are greatly improved.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a kit for jointly detecting the contents of aldehyde and ketone reductase 1B10 (AKR1B10) and alpha-fetoprotein (AFP) by time-resolved fluorescent immunoassay, which can be used for liver cancer, Screening, diagnosis, curative effect judgment, prognosis assessment or recurrence monitoring of hepatitis, cirrhosis and other liver diseases. Background technique [0002] Alpha-fetoprotein (α-fetoprotein, αFP or AFP) is a carcinoembryonic glycoprotein with a relative molecular mass of about 68kDa, which is synthesized by the fetal liver and yolk sac. In adults, AFP is produced by liver cells, and its serum content is very small, but when the liver cells become cancerous, they resume the function of producing AFP, and as the disease progresses, the AFP content in serum will increase sharply. Detection of serum AFP level has extremely high clinical value for early diagnosis of prima...

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): G01N33/68G01N33/577G01N33/576G01N33/574G01N33/573
CPCG01N33/6893G01N33/577G01N33/57488G01N33/57476G01N33/57438G01N33/576G01N33/573G01N2333/90203G01N2333/471G01N2800/085G01N2800/52
Inventor 曹哲陈志勇祝跃球
Owner 湖南莱拓福生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com