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A high-throughput combined detection reagent for hepatitis C virus antigen antibody

A hepatitis C virus, combined detection technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of missed diagnosis, obtaining test results, strong technical ability, etc., to achieve enhanced sensitivity, guaranteed specificity, and high application value. Effect

Active Publication Date: 2017-01-25
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing HCV clinical detection methods mainly include PCR detection, hepatitis C virus antibody detection and hepatitis C virus core antigen detection kit. Among the three detection methods, the PCR detection method has high sensitivity and accurate results, but the method is cumbersome to operate , the required personnel have strong technical skills, and it is not easy to promote it in various hospitals; the hepatitis C virus antibody detection reagent is the most common hepatitis C detection method in clinical practice, but there is a "window period" for this detection, that is, in the hepatitis C infection In the early stage, the immune antibody did not appear in the patient's body, and the test result could not be obtained in the early stage, resulting in a missed diagnosis; the hepatitis C virus core antigen detection reagent has gradually been accepted by hospitals in recent years, and its market share has also continued to increase , but for the follow-up detection of hepatitis C patients, there is no good tracking effect

Method used

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  • A high-throughput combined detection reagent for hepatitis C virus antigen antibody
  • A high-throughput combined detection reagent for hepatitis C virus antigen antibody
  • A high-throughput combined detection reagent for hepatitis C virus antigen antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] The components and concentrations of the detection reagents were

[0070] Component 1 (R1) FITC fluorescent microsphere reagent:

[0071] FITC Highlight Microspheres Coated with HCV Antibody 0.1%

[0072] FITC low brightness microspheres coated with HCV protein 0.1%

[0073] Tris 12.1g / L

[0074] BSA 0.5g / L

[0075] PC-300 0.1%

[0076] Prepare with deionized water and adjust the pH to 7.6 with HCl.

[0077] Component 2 (R2) PE labeling reagent:

[0078] PE-labeled HCV-cAg antibody 0.1%

[0079] PE-labeled NS3 protein 0.1%

[0080] PE-labeled NS4 protein 0.1%

[0081] PE-labeled NS5 protein 0.1%

[0082] Tris 12.1g / L

[0083] BSA 0.5g / L

[0084] PC-300 0.1%

[0085] Prepare with deionized water and adjust the pH to 7.6 with HCl.

[0086] Cleaning fluid:

[0087] Disodium hydrogen phosphate 1.974g / L

[0088] Potassium dihydrogen phosphate 0.224g / L

[0089] Sodium chloride 0.9g / L

[0090] Tween-20 0.5%

[0091] All components were dissolved in deionized ...

Embodiment 2

[0113] The components and concentrations of the detection reagents were

[0114] Component 1 (R1) FITC fluorescent microsphere reagent:

[0115] FITC highlight microspheres coated with HCV core antigen antibody 0.5%

[0116] FITC low brightness microspheres coated with HCV protein 0.5%

[0117] Tris 12.1g / L

[0118] BSA 0.5g / L

[0119] PC-300 0.1%

[0120] Prepare with deionized water and adjust the pH to 7.6 with HCl.

[0121] Component 2 (R2) PE labeling reagent:

[0122] PE-labeled HCV-cAg antibody 0.5%

[0123] PE-labeled NS3 protein 0.5%

[0124] PE-labeled NS4 protein 0.5%

[0125] PE-labeled NS5 protein 0.5%

[0126] Tris 12.1g / L

[0127] BSA 0.5g / L

[0128] PC-300 0.1%

[0129] Prepare with deionized water and adjust the pH to 7.6 with HCl.

[0130] Cleaning solution, positive control, negative control and detection method are as in Example 1.

Embodiment 3

[0132] The components and concentrations of the detection reagents were

[0133] Component 1 (R1) FITC fluorescent microsphere reagent:

[0134] FITC Highlight Microspheres Coated with Hepatitis C Core Antibody 1%

[0135]FITC low brightness microspheres coated with HCV protein 1%

[0136] Tris 12.1g / L

[0137] BSA 0.5g / L

[0138] PC-300 0.1%

[0139] Prepare with deionized water and adjust the pH to 7.6 with HCl.

[0140] Component 2 (R2) PE labeling reagent:

[0141] PE-labeled HCV-cAg antibody 1%

[0142] PE-labeled NS3 protein 1%

[0143] PE-labeled NS4 protein 1%

[0144] PE-labeled NS5 protein 1%

[0145] Tris 12.1g / L

[0146] BSA 0.5g / L

[0147] PC-300 0.1%

[0148] Prepare with deionized water and adjust the pH to 7.6 with HCl.

[0149] Cleaning solution, positive control, negative control and detection method are as in Example 1.

[0150] Effect test 1:

[0151] Analyze the accuracy of the kit for combined detection of hepatitis C virus antigen-antibody...

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Abstract

Disclosed is a reagent for high throughput combined detection of a hepatitis C virus antigen and antibody. Using the flow cytometry principle, different concentrations of FITC fluorescein-labeled microspheres correspond to coated antibodies and antigens; after addition of a detection sample, the microspheres, the detection sample and a PE-labeled paired antibody and antigen form a sandwich structure; and under the action of 480nm exciting light, the effects of simultaneous detection of a hepatitis C virus antigen and antibody can be achieved in a high throughput way according to different emitted light intensities of fluoresceins at different concentrations. By means of the utilization of the fluorescent coloration effect, the reaction sensitivity is enhanced, and a hepatitis C virus antibody and core antigen can be detected simultaneously by the means of utilization of FITC-labeled microspheres for simultaneous detection of the antigen and antibody.

Description

technical field [0001] The invention belongs to the technical field of hepatitis C detection, in particular to a high-throughput combined detection reagent for hepatitis C virus antigen-antibody. Background technique [0002] Hepatitis C is a global infectious disease caused by hepatitis C virus (Hepatitis C virus, HCV) infection. It is estimated that there are currently 170 million hepatitis C virus-infected people in the world, and the hepatitis C infection rate in my country is about 30%. At present, there are at least 40-60 million hepatitis C patients. Among them, 80-85% of infected patients will develop chronic hepatitis C, and 20% of them will develop liver fibrosis, and finally 4-5% of patients with liver fibrosis will develop hepatocellular carcinoma, which is very harmful. [0003] At present, there is no specific drug for the treatment of hepatitis C virus, and the development of hepatitis C vaccine is also difficult due to the rapid mutation of HCV, and it is di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/545G01N33/533
CPCG01N21/64G01N33/533G01N33/569G01N33/68
Inventor 甘宜梧王进谭柏清李静王绮
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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