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Nondiagnostic HCV (hepatitis c virus) antibody immunodetection method and kit

An immune detection method and immune detection technology, applied in the field of detection of HCV antibodies, can solve problems such as the detection of low-concentration HCV antibody levels in the early stage of infection, and achieve the effects of easy automation, low toxicity, and simple operation

Inactive Publication Date: 2017-05-31
THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the lower detection limit of ELISA is still limited, and it cannot meet the detection of low concentration HCV antibody levels in the early stage of infection.

Method used

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  • Nondiagnostic HCV (hepatitis c virus) antibody immunodetection method and kit
  • Nondiagnostic HCV (hepatitis c virus) antibody immunodetection method and kit
  • Nondiagnostic HCV (hepatitis c virus) antibody immunodetection method and kit

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Embodiment 1

[0052] The various solution formulations used in the embodiment are as follows:

[0053] Sodium phosphate-sodium chloride buffer solution (SPSC) (50mM Na 2 HPO 4 / 1.0M NaCl): 17.9g of Na 2 HPO 4 ·12Biotin-H2O, 58.5g of NaCl and a little ultrapure water to dissolve, adjust the pH to 7.5 with hydrochloric acid, and make up to 100mL.

[0054] 10mM phosphate buffer solution (PBS) (pH=7.4): 8g NaCl, 0.2g KCl, 1.44g Na 2 HPO 4 12Biotin-H2O, 0.24g KBiotin-H2PO 4 Dissolve in 1L ultrapure water, adjust pH=7.4 with hydrochloric acid, and store at 4°C after autoclaving.

[0055] Preparation of Biotin-I: 3.3 nmol Biotin-I synthesized according to the known sequence was dissolved in 33 μl ultrapure water to 100 μM and diluted to 10 nM with SPSC buffer. Divide and store at -20°C for later use. Avoid repeated freezing and thawing before use.

[0056] Preparation of H1 and Biotin-H2: Dissolve 2.1nmol H1 synthesized according to the known sequence in 21μl ultrapure water, dissolve 2.2n...

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Abstract

The invention relates to a nondiagnostic HCV (hepatitis c virus) antibody immunodetection method and a kit. The method comprises steps as follows: an HCV antigen specifically captures an HCV antibody in a detection sample, a biotin-labeled second antibody is added, and an antigen-antibody-biotin-labeled second antibody compound is formed after incubation; in the presence of streptavidin, a biotin primer Biotin-I is added, incubation is performed, so that the compound and the Biotin-I are bound, and then, a hair grip chain H1 and a hairpin chain Biotin-H2 are added for a hybridization reaction; avidin-labeled horseradish peroxidase is added, a substrate solution is catalyzed to develop, and quantitative detection of the HCV antibody is performed by measuring the absorbance. ELISA (enzyme-linked immuno sorbent assay) is introduced into an HCR (hybridization chain reaction), HCR is used for signal amplification, ultra-sensitive detection of the HCV antibody is realized, and the lower limit of detection is lower than 10 pg / mL and is improved by at least two orders of magnitude as compared with conventional ELISA.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and in particular relates to a non-diagnostic method for detecting HCV antibodies and a kit. Background technique [0002] Hepatitis C virus (HCV) is a single-stranded positive-stranded RNA virus belonging to the Flaviviridae family of hepadnaviruses. HCV is transmitted through blood, and infection in humans can cause hepatitis, and it is easy to develop chronic hepatitis, liver cirrhosis, and even liver cancer. According to the World Health Organization report, about 130 million to 150 million people in the world are infected with HCV virus, with 3 to 4 million new cases of virus infection every year, and about 500,000 people die of liver disease related to HCV infection. HCV has become a serious threat to human health. of public health issues. Therefore, it is necessary to screen some populations, especially high-risk populations, for HCV antibodies. Selecting a sensitive, accurate, time...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/576
CPCG01N33/56983G01N33/5767
Inventor 桂嵘聂新民黄蓉江静李建
Owner THE THIRD XIANGYA HOSPITAL OF CENT SOUTH UNIV
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