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HCV viral antigen, ligand tube-type PCR detection kit, preparation and use thereof

A technology for detecting kits and virus antigens, which is applied in biochemical equipment and methods, measuring devices, and microbial determination/inspection. , to achieve high sensitivity and specificity

Inactive Publication Date: 2009-08-12
兰州普利生物技术开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the sandwich method uses a traditional enzyme-linked detection antibody to detect the capture molecule-antigen complex, and then uses the reporter enzyme molecule to label the oligonucleotide, which requires chemical synthesis steps and additional labor, and the primers used for labeling cannot resolve the impurities. problems and primer-dimer amplification problems, therefore, cannot solve the problem of quantitative detection

Method used

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  • HCV viral antigen, ligand tube-type PCR detection kit, preparation and use thereof
  • HCV viral antigen, ligand tube-type PCR detection kit, preparation and use thereof
  • HCV viral antigen, ligand tube-type PCR detection kit, preparation and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0081] Example 1: Novel Immuno-PCR Detection

[0082] Novel PCR detection process (see figure 2 ) refers to: after the PCR tube is treated, it is first coated with an antibody, and then incubated with the test sample to form an antibody-antigen complex; The unbound detection antibody is finally subjected to PCR amplification and analysis and processing of the amplified product for the ligand-modified sequence. The specific steps are as follows:

[0083] 1. Preparation of PCR tube: Treat a 0.2ml polystyrene PCR tube with 50ul of glutaraldehyde with a volume concentration of 1-20% at 37±1°C for 1-3 hours; Rinse the PCR tube three times every minute; coat the monoclonal antibody 2ug / ml capturing HCV virus antigen on the PCR tube with sodium bicarbonate buffer solution with a pH value of 9.6, and then use 100ul of the PCR tube with a pH value of 7.4 containing 5 The sodium bicarbonate buffer of % skimmed milk powder, glycine, bovine serum albumin and tortoise sperm DNA was tre...

Embodiment 2

[0103] Embodiment 2: Ligand detection

[0104] Ligand detection process (see image 3 ) refers to: after the PCR tube is treated, it is first coated with the ligand that recognizes the ligand, and then incubated with the detection sample to form a ligand-ligand complex; after washing, the recognition and detection ligand labeled with the modified sequence is added. After co-incubating with different ligands of the body, the unbound detection ligands were washed away, and finally PCR amplification and amplification products were analyzed and processed for the ligand-modified sequences. The specific steps are as follows:

[0105] 1. Preparation of PCR tubes: Treat 0.2ml polystyrene PCR tubes with 50ul, 1-20% glutaraldehyde at 37±1°C for 1-3 hours; wash the PCR tubes three times with ultrapure water for 2-5 minutes each time After 50ul streptavidin (streptavidin) is coated on the PCR tube, use 60ul, the sodium bicarbonate buffer that pH value is 7.4 to contain 5% skimmed milk p...

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Abstract

The invention relates to a PCR tubular detection kit for detecting an HCV virus antigen and a ligand thereof. The kit comprises a PCR tube of an antibody or a petunidin of a sexual human immunodeficiency virus (HCV) target molecule which is obtained by capture, detection agents of A solution and B solution for detecting a ligand or an antigen of the sexual human immunodeficiency virus (HCV), a standard product and a reference product. The invention converts a ligand signal into a nucleic acid petunidin signal by direct combination of specific oligonucleotide petunidin (specific oligonucleotide petunidin carrying signal molecules) and the ligand, and then measures the ligand by PCR augmentation for amplification and detection, so the detection kit has the characteristics for detecting the ligand with quickness, high sensitivity and quantitive measurement. The invention also discloses a preparation method for the kit and an application method of the kit for detecting sexual human immunodeficiency virus (HCV) in the specific oligonucleotide ligand.

Description

technical field [0001] The invention relates to a detection kit, in particular to a tubular PCR detection kit for HCV virus antigen and ligand, a preparation method and application thereof. Background technique [0002] Early protein detection is based on the low dissociation constant and certain specificity of the antibody and the specific protein to be tested, and adopts a simple and convenient screening method - antibody capture method. This method is to detect whether there is an antigen in the sample: firstly, the antigen is coated on a solid support, then the antibody is used to bind the antigen to form a complex, the unbound antibody is washed away, and finally the labeled molecule that is specifically recognized by the bound antibody is used for detection. Binding to antibodies; antigens and antibodies can also react to form complexes first, then bind to solid supports, and then detect the complexes. Many antibody capture methods use indirect methods to detect antib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N33/577G01N21/00
Inventor 廖世奇
Owner 兰州普利生物技术开发有限公司
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