Immunological marker detection method, reagent, and detection kit

A marker and anti-immune technology, which is applied in the field of medical testing, can solve problems such as difficult automation, and achieve the effects of improving detection efficiency, high accuracy, and simplifying operation steps

Inactive Publication Date: 2016-12-14
FUJIAN COSUNTER PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invension describes methods used for detecting biomarkers (biopolymer) on blood samples or other materials like milk by combining them together into small pieces called magnetized microparticles. These tiny objects have unique properties such as being able to attach themselves easily onto any material they come across during testing without affecting their original function. This makes these techniques simpler than previously possible but also more efficient at identifying new ones from existing substances.

Problems solved by technology

This patents describes various technical means related to improving the accuracy and reliability of measuring certain types of materials called autoimmunity markers. These technologies involve analyzing samples containing both weakly bound material like protein(protein), nucle acid, DNA, amino acids, peptide, sugar chains, nucleations, cells, blood clots, tumour necroid bodies, bacteria, viruses, liver spikes, cancerous growths, and others. By doing this analysis, we aim to develop effective therapies against all kinds of illnesses caused by these agents.

Method used

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  • Immunological marker detection method, reagent, and detection kit
  • Immunological marker detection method, reagent, and detection kit
  • Immunological marker detection method, reagent, and detection kit

Examples

Experimental program
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Embodiment 1

[0089] Example 1. Preparation of DCP Magnetic Particle Separation Immunofluorescence Analysis Detection Reagent

[0090] 1. Preparation of DCP antigen

[0091] (1) Construction of pColdⅡ-DCP expression vector

[0092] Design primers and synthesize the DCP gene with reference to the gene sequence of human DCP in GenBank. After PCR amplification, 1% agarose gel electrophoresis was used to detect the DCP amplification product, and the target band was excised, and the target fragment was recovered using a gel recovery kit. The prokaryotic expression vector pColdⅡ was ligated with restriction endonucleases NdeⅠ and HindⅢ, transformed into E. coli E.coli.DH5α for amplification, and then transferred into E.coli.BL21 (DE3) competent cells, inoculated In LB solid medium containing 100 mg / L ampicillin (Amp), the plasmid was extracted, identified by NdeI and HindIII enzyme digestion and sequenced. The recombinant plasmid pColdⅡ-DCP was double digested with 5'NdeⅠand 3'HindⅢ, and two sp...

Embodiment 2

[0149] Example 2. Preparation of AFP-L3 Magnetic Particle Separation Immunofluorescence Analysis Detection Reagent

[0150] 1. Preparation of AFP-L3 antigen

[0151] (1) Construction of pColdⅡ-AFP-L3 expression vector

[0152] Refer to the gene sequence of human AFP-L3 in GenBank to design primers and synthesize the AFP-L3 gene. After PCR amplification, 1% agarose gel electrophoresis was used to detect the AFP-L3 amplification product. Recycle the target fragment. The prokaryotic expression vector pColdⅡ was ligated with restriction endonucleases NdeⅠ and HindⅢ, transformed into E. coli E.coli.DH5α for amplification, and then transferred into E.coli.BL21 (DE3) competent cells, inoculated In LB solid medium containing 100 mg / L ampicillin (Amp), the plasmid was extracted, identified by NdeI and HindIII enzyme digestion and sequenced. The recombinant plasmid pColdⅡ-AFP-L3 was double digested with 5'NdeⅠ and 3'HindⅢ, and two specific bands appeared, whose positions were consist...

Embodiment 3

[0195] Example 3. Preparation of GP73 Magnetic Particle Separation Immunofluorescence Analysis Detection Reagent

[0196] 1. Preparation of GP73 antigen

[0197] (1) Construction of pColdⅡ-GP73 expression vector

[0198] Design primers with reference to the human GP73 gene sequence in GenBank and synthesize the GP73 gene. After PCR amplification, 1% agarose gel electrophoresis was used to detect the GP73 amplification product, and the target band was excised, and the target fragment was recovered using a gel recovery kit. The prokaryotic expression vector pColdⅡ was ligated with restriction endonucleases NdeⅠ and HindⅢ, transformed into E. coli E.coli.DH5α for amplification, and then transferred into E.coli.BL21 (DE3) competent cells, inoculated In LB solid medium containing 100 mg / L ampicillin (Amp), the plasmid was extracted, identified by NdeI and HindIII enzyme digestion and sequenced. The recombinant plasmid pColdⅡ-GP73 was digested with 5'NdeⅠand 3'HindⅢ, and two speci...

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Abstract

The invention discloses an immunological marker detection method, a used reagent, and a detection kit. The detection method comprises the following steps: step one, preparing a magnetic micro particle (A) of a surface coupling anti-immunological marker antibody and a magnetic micro particle (B) of a surface coupling immunological marker antigen; step two, preparing a fluorescence labeled anti-immunological marker antibody; and step three, detecting the immunological marker content: reacting a blood plasma or serum sample with the magnetic micro particle (A), after reaction, eluting the immunological marker, adding the fluorescence labeled anti-immunological marker antibody, carrying out incubation, then adding the magnetic micro particle (B) , keeping on incubation, carrying out magnetic separation, detecting the fluorescence value of the sample by a fluorospectro photometer, and calculating the immunological marker content of the sample.

Description

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Claims

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Application Information

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Owner FUJIAN COSUNTER PHARMA
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