Protein marker for simultaneously detecting specificity of multiple diseases in blood serum and kit thereof

A technology of protein markers and kits, which is applied in the field of protein markers and kits, can solve the problems of complex, time-consuming, and large sample volume in the pre-processing of sputum samples

Inactive Publication Date: 2011-09-14
薛芳 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When performing enzyme-linked immunoassay, each experiment takes about one and a half to two hours, and tuberculosis sputum culture takes up to six weeks. When performing these experiments, each experiment requires an exp

Method used

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  • Protein marker for simultaneously detecting specificity of multiple diseases in blood serum and kit thereof
  • Protein marker for simultaneously detecting specificity of multiple diseases in blood serum and kit thereof
  • Protein marker for simultaneously detecting specificity of multiple diseases in blood serum and kit thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0121] At the same time, the extraction of protein markers specific to hepatitis A, hepatitis B, hepatitis C, AIDS, syphilis, and tuberculosis in serum is detected.

[0122] Instruments and reagents used: nano-magnetic beads with carboxyl as a ligand: purchased from Bruker Company, product number MBWCX; lysate: 9M Urea, mass ratio of 2% CHAPS, 50mM Tris-HCl, pH9.0 purchased from SIGMA Company , product number U0631, C5070, T2819; buffer: 50mM NaAC, pH4.0 purchased from SIGMA Company, product number S7545; deionized water: purchased from SIGMA Company, product number 27,073-3; sinapinic acid: purchased from SIGMA Company, product number S5019; Trifluoroacetic acid (TFA): purchased from SIGMA, product number T6508; protein reader: purchased from BIORAD company, product number PBSIIc; quality control serum: purchased from BIORAD company, product number StandardS10.

[0123] (1) Serum samples: A total of 230 serum samples were collected from Liaoning International Travel Health Ca...

Embodiment 2

[0135] Simultaneous detection of multiple diseases in serum The composition of the kit:

[0136] 1) Nano magnetic beads with carboxyl as ligand;

[0137] 2) Lysis solution: 9M Urea, 2% CHAPS, 50mM Tris-HCl, pH9.0;

[0138] 3) Buffer: 50mM NaAC, pH4.0;

[0139] 4) Deionized water;

[0140] 5) sinapic acid;

[0141] 6) Trifluoroacetic acid;

[0142] 7) Protein reader;

[0143] 8) Quality control serum.

Embodiment 3

[0145] Simultaneous clinical trial of rapid detection of infectious disease protein marker kit.

[0146] (1) Serum samples: 120 cases of infectious diseases (20 cases each of hepatitis A, hepatitis B, hepatitis C, AIDS, syphilis and tuberculosis) and 30 normal healthy samples.

[0147] (2) Collection of serum samples: Collect 5 mL of whole blood from the patient on an empty stomach in the morning, place it in a vacuum negative pressure tube without anticoagulant, let it stand at room temperature for 1 hour, centrifuge at 2000g for 20 minutes, and take the supernatant. Aliquot into 50μl / tube and store in -80℃ refrigerator.

[0148] (3) Sample preparation: Take out the serum from the -80°C refrigerator and put it on ice to thaw. Centrifuge at 10000rpm for 2min. 10 μl of the supernatant was mixed with 20 μl of U9 lysate (9 mol / L Urea, 20 g / L CHAPS, 10 g / L DTT, 50 mmol Tris-HCl, pH 9.0). After shaking and incubating at 4°C for 30 min, 370 μl of sodium acetate buffer (100 mmol / L s...

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Abstract

The invention discloses a group of protein marker for simultaneously detecting the specificity of hepatitis A, hepatitis B, hepatitis C, Aids, syphilis and tuberculosis in blood serum, a kit for detecting the group of specific proteins and application thereof. The protein marker for detection the specificity in the invention comprises seven proteins with different molecular weights in a serum proteome map: A: 5232.13, B: 8784.25, C: 2017.5, D: 4309.19, E 5489.97, F: 8076.62 and G: 2155.92. A detection method for the specific proteins comprises the steps of: processing a serum sample to be detected by using a lysis solution; incubating and capturing serum proteins by using a nano magnetic bead with carboxyl as a genin; and eluting the nano magnetic bead, and detecting seven specific proteins in eluted ingredients by using a protein reader. The kit and the detection method in the invention have the advantages of rapidness, sensitivity, specificity, flexibility, simplicity for operation and the like.

Description

technical field [0001] The present invention relates to the field of biotechnology, and specifically refers to a simultaneous detection of multiple disease-specific protein markers in serum and a kit thereof. The various diseases described in this application are hepatitis nails, hepatitis B, hepatitis C, AIDS, syphilis, and tuberculosis . Background technique [0002] With the completion of the Human Genome Project, life science has ushered in the challenge of the post-genome era - Human Proteomics (Proteome). Because although the human genome contains all the genetic information of our body, it is only a blueprint for making proteins. Proteins make up cells and perform various life phenomena through their physiological functions. Therefore, only protein is the performer and bearer of life phenomena. Protein is also the determinant of the differences between various types of cells in the body. Although all cells have the same genome, different genes are open (expressed) ...

Claims

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Application Information

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IPC IPC(8): G01N27/64
Inventor 薛芳李云峰李潇蔡鹏毕秀欣
Owner 薛芳
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