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Hepatitis C virus antibody rapid test strips

A hepatitis C virus and test strip technology, applied in the field of biopharmaceuticals, can solve the problems of unknown overall sensitivity, complex process, low detection sensitivity, etc., and achieve rapid screening, simplified operation process, and high overall coincidence rate. Effect

Active Publication Date: 2021-06-29
BEIJING KAWIN TECH SHARE HLDG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent CN103575884A discloses a rapid diagnostic kit for hepatitis C virus by colloidal gold method. The hepatitis C virus antigen coated on the nitrocellulose membrane detection line contains six proteins of core, E1, E2, NS3, NS4, and NS5. The six genes are all constructed on the vector and expressed in the transformed host bacteria, the process is relatively complicated, and the detection sensitivity is not high
CN102109523A discloses a hepatitis C IgG, IgM antibody joint detection kit, the hepatitis C virus antigen coated on the nitrocellulose membrane detection line includes IgM antibody and HCV multi-epitope composite antigen (including core protein 1-16aa, 132-141aa , E1 protein 126-135aa, NS3 protein 139-147aa and NS5768-775aa, multi-epitope complex antigen contains 55 amino acids), the overall sensitivity is unknown

Method used

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  • Hepatitis C virus antibody rapid test strips
  • Hepatitis C virus antibody rapid test strips
  • Hepatitis C virus antibody rapid test strips

Examples

Experimental program
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Effect test

Embodiment 1

[0023] The preparation of embodiment 1 colloidal gold solution

[0024] Heat 50ml of 0.01% (weight / volume) chloroauric acid to boiling, then add 0.5ml of 1% trisodium citrate solution, the reaction solution turns from off-white to deep purple at first, and continue to boil for 3 minutes to obtain the product.

Embodiment 2

[0025] The preparation of embodiment 2 gold standard pad

[0026] Take Protein G (product of Arista Biologicals), dialyze the Protein G solution against 0.005M pH7.0 PBS solution overnight at 4°C to remove excess salt ions, then centrifuge at 100,000g at 4°C for 1h, and take the supernatant, and use the BCA method to detect the protein Concentration, and diluted to 2mg / ml with 0.005M PBS solution of pH7.0.

[0027] Colloidal gold solution with 0.1 M K 2 CO 3 or 0.1M HCl to adjust the pH to 9.0, take 50ml of Protein G solution, add 2ml of colloidal gold solution under electromagnetic stirring, stir for 5min, add 1ml of 5% BSA solution, continue stirring for 10min, then centrifuge at 10000g for 5min, carefully suck off the For the supernatant, the pellet was resuspended in 50 ml of PBS buffer (pH 7.0) containing 0.2 mg / ml PEG200000 and 5 mg / ml sodium azide, and stored at 4°C.

[0028] Spread the colloidal gold-labeled Protein G solution evenly on the glass fiber membrane on a...

Embodiment 3

[0029] Coating of embodiment 3 detection line (T line) and quality control line (C line)

[0030] Preparation of recombinant HCV multi-epitope composite antigen

[0031] Through PCR amplification technology, the gene encoding the amino acid sequence of SEQ ID NO: 1 was constructed into the pET-his vector (from the Addgene website), and then transformed into BL21 (DE3) bacteria, under the induction of IPTG, the protein expression was performed, and the expression The protein was purified by Ni column of GE Company, and the purified protein was purified by high-performance liquid chromatography (HPLC) and SDS-PAGE to obtain a conforming antigen with a purity of more than 95%, which was preserved.

[0032] Goat anti-human IgG antibody: purchased from Abcam.

[0033] Coating buffer: 0.05M PB, 0.02% sucrose, 0.1% Azide, pH 7.4.

[0034] Coating of T line: take the recombinant HCV multi-epitope composite antigen, dilute it to 0.8 mg / ml with coating buffer, and streak at 1 μl / cm. ...

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Abstract

This product is a test strip for the rapid detection of hepatitis C virus antibody saliva. The test strip is composed of a bottom plate and a casing. Colloidal gold-labeled Protein G, nitrocellulose membrane coated with recombinant antigen specifically binding to hepatitis C virus antibody (test line), and a goat anti-human IgG (quality control line). The rapid detection test strip has the advantages of good conservation and strong specificity, can detect infection by hepatitis C virus of all genotypes (types 1-6), and is not interfered by hepatitis B virus and the like.

Description

technical field [0001] The invention relates to a test strip for detecting whether there is hepatitis C virus antibody in saliva, urine and / or blood, and belongs to the technical field of biopharmaceuticals. Background technique [0002] Hepatitis C virus (HCV) infection is the most common chronic blood-borne infection in the United States. Although the number of new infections has declined, the burden of chronic infection is still substantial. According to the World Health Organization, the global HCV infection rate is about 2.8%. It is estimated that about 185 million people are infected with HCV, and the number of deaths caused by HCV infection every year There are about 350,000 cases. Hepatitis C can lead to chronic inflammation, necrosis and fibrosis of the liver, and some patients can develop liver cirrhosis or even hepatocellular carcinoma. Currently there is no effective preventive hepatitis C vaccine available, and the mortality rate (death caused by liver failure...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/576G01N33/68G01N33/58G01N33/532
CPCG01N33/532G01N33/576G01N33/58G01N33/6854G01N2800/26
Inventor 黎维
Owner BEIJING KAWIN TECH SHARE HLDG
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