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A kind of hepatitis C virus antigen-antibody combined detection kit and preparation method thereof

A hepatitis C virus, antigen antibody technology, applied in the field of immunoanalysis medicine, can solve the problems of simultaneous detection, low antibody sensitivity, and poor specificity, and achieve the effects of shortening the window period, strong detection specificity, and low cost

Active Publication Date: 2016-09-28
BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problems that the sensitivity of HCV antigen and antibody detection is not high, the specificity is poor, and cannot be detected simultaneously in the prior art, the technical scheme adopted in the present invention is:

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Reagent test kit

[0032] A combined detection kit for hepatitis C virus antigen and antibody, comprising:

[0033] (1) Calibrator: a group of substances used to make a standard curve, 6 bottles;

[0034] (2) Double-labeled enzyme conjugates;

[0035] (3) Negative and positive controls;

[0036] (4) Luminescence liquid; said luminescence liquid comprises luminescence liquid 1 and luminescence liquid 2, and luminescence liquid 1 contains 0.7g / L luminol, 0.9g / L cinnamic acid, 0.2g / L 4-iodophenylboronic acid, 0.25g / L p-iodophenol, 25ml / L dimethylformamide, 5g / L polyvinyl alcohol,

[0037] 8g / L polyvinylpyrrolidone, 3g / L polyethylene glycol 600, 4g / L ethylenediaminetetraacetic acid, 1.6 million units / L gentamicin sulfate, 0.4g / L carbamide peroxide, 0.1mol of pH9.0 / L Tris buffer solution; luminescence solution 2 is 0.1mol / L Tris buffer solution containing 0.1mg / ml acridinium ester derivative, 3g / L polyethylene glycol 600, 0.1% TWEEN-20pH9.0;

[0038] (5) Microwell coa...

Embodiment 2

[0040] The preparation of kit described in embodiment 1

[0041] (1) The preparation method of the double-labeled enzyme conjugate is: use the improved sodium periodate oxidation method to label horseradish peroxidase on the HCV chimeric antigen, and use the EDC method to label alkaline phosphatase to another HCV For the monoclonal antibody, dilute the chimeric antigen-HRP at 0.1 μg / ml, and HCV mab-ALP at 0.2 μg / ml into the enzyme conjugate diluent containing bovine serum albumin and proclin300, as the enzyme working solution, in 2~ Store at 8°C.

[0042] (2) Negative and positive controls are respectively: use negative mixed human serum as negative control, add HCV-1 antibody in negative serum as positive control 1, add HCV-2 antibody in negative serum as positive control 2, add HCV-2 antibody in negative serum as positive control 2, HCV antigen was added as positive control 3.

[0043](3) The preparation method of the microwell coated plate is: dilute the chimeric antigen ...

Embodiment 3

[0045] The preparation of kit described in embodiment 1

[0046] (1) The preparation method of the double-labeled enzyme conjugate is: use the improved sodium periodate oxidation method to label horseradish peroxidase on the HCV chimeric antigen, and use the EDC method to label alkaline phosphatase to another HCV For the monoclonal antibody, dilute the chimeric antigen-HRP at 0.1 μg / ml, and HCV mab-ALP at 0.2 μg / ml into the enzyme conjugate diluent containing bovine serum albumin and proclin300, as the enzyme working solution, in 2~ Store at 8°C.

[0047] (2) Negative and positive controls are respectively: use negative mixed human serum as negative control, add HCV-1 antibody in negative serum as positive control 1, add HCV-2 antibody in negative serum as positive control 2, add HCV-2 antibody in negative serum as positive control 2, HCV antigen was added as positive control 3.

[0048] (3) The preparation method of the microwell coated plate is: dilute the chimeric antigen...

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Abstract

A hepatitis c virus antigen-antibody joint detection reagent box is characterized by comprising a calibrator(1), a double-marker enzyme conjugate (2), a negative and positive contrast (3), light-emitting liquid (4) and a micropore coated plate (5), wherein the light-emitting liquid contains light-emitting liquid 1 and light-emitting liquid 2, the light-emitting liquid 1 contains luminal 0.7 g / L, cinnamic acid 0.9 g / L, 4-iodophenylboronic acid 0.2 g / L, iodobiphenol 0.25 g / L, dimethylformamide 25 ml / L, polyving akohol 5 g / L, polyvinylpyrrolidone 8 g / L, polyethylene glycol 600 3 g / L, ethylenediamine tetraacetic acid 4 g / L, gentamicin sulfate 1600 thousands / L, urea peroxide 0.4 g / L, and pH 9.0 Tris buffer solution 0.1 mol / L. The light-emitting liquid 2 contains acridinium ester derivative 0.1 mg / ml, polyethylene glycol 600 3 g / L and 0.1 mol / L of pH 9.0 Tris buffer solution containing 0.1% of TWEEN-20. The invention further discloses a preparation method and a using method of the reagent box. The hepatitis c virus antigen-antibody joint detection reagent box has the advantages of being quick in reaction and low in cost.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, specifically, the invention provides a hepatitis C virus (HCV) antigen-antibody combined detection kit and a preparation method thereof. Background technique [0002] Hepatitis C virus (Hepatitis C virus, HCV) is a class of single-strand positive-strand RNA virus, which has an envelope, a thorn-like protrusion structure on the surface, and large variations in virus particles. Blood transfusion and injection are the important transmission routes of HCV in my country. The development of hepatitis C is hidden, the clinical manifestations are not typical, and the clinical diagnosis and curative effect observation depend on the laboratory test results. [0003] The hepatitis C virus genome contains an open reading frame (ORF), which can encode a viral precursor polypeptide of amino acid residues, which respectively constitute the core protein (c), matrix protein (M), coat protein (E) and Anothe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/56983G01N33/5767
Inventor 刘萍栾大伟张振斌陈露鸾侯玉文杨桂霞李克锦王东生曾小莉
Owner BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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