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Reagent for high throughput combined detection of 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, strong technical ability, complicated method operation, etc., to achieve the effect of enhanced sensitivity and high application value

Active Publication Date: 2015-10-28
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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  • Reagent for high throughput combined detection of hepatitis c virus antigen-antibody
  • Reagent for high throughput combined detection of hepatitis c virus antigen-antibody
  • Reagent for high throughput combined detection of hepatitis c virus antigen-antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[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 deioni...

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-an...

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Abstract

The invention provides a reagent for high throughput combined detection of hepatitis c virus antigen-antibody. By the flow cytometry theory, different concentrations of FITC fluorescein-labeled microspheres are correspondingly coated with antibody and antigen; after addition of a detection sample, the microspheres, the detection sample and the PE-labeled paired antibody and antigen form a sandwich structure; and under the action of 480nm exciting light, the effects of simultaneous detection of hepatitis c virus antigen-antibody can be achieved in a high throughput way according to different emitted light intensities of fluoresceins with different concentrations. By the utilization of fluorescent color-developing effect, reaction sensitivity is enhanced, and hepatitis c virus antibody and core antigen can be detected simultaneously by the utilization of FITC-labeled microspheres for simultaneous detection of antigen-antibody. The reagent is more accurate in hepatitis c detection, has no defect in detection leakage, achieves the early detection effect and has high application value in prevention of hepatitis c.

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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/545G01N33/533
CPCG01N21/64G01N33/533G01N33/569G01N33/68G01N33/6854G01N33/54313G01N33/545G01N33/56983
Inventor 甘宜梧王进谭柏清李静王绮
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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