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Kit and protein chip for diagnosis of tuberculosis latent infection

A protein chip and kit technology, applied in the field of biomedicine, can solve problems such as limited efficiency, high false positive rate, and trouble, and achieve high sensitivity and specificity, and low sample demand.

Active Publication Date: 2020-05-12
RAYBIOTECH INC GUANGZHOU +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

PPD skin test is relatively simple, but it has cross-reaction with BCG, and the false positive rate is relatively high, especially in the case of widespread vaccination of BCG in China, its use efficiency is obviously limited
IGRA technology detects the expression of tuberculosis-specific IFNg. Although the specificity is significantly improved, the current method is relatively laborious and time-consuming, and the price is relatively expensive, which seriously limits the clinical promotion and use.

Method used

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  • Kit and protein chip for diagnosis of tuberculosis latent infection
  • Kit and protein chip for diagnosis of tuberculosis latent infection
  • Kit and protein chip for diagnosis of tuberculosis latent infection

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0033] A kind of preparation method of the protein chip that is used for the diagnosis of tuberculosis latent infection of the present invention comprises the following steps:

[0034] Immobilize the capture antibody on the substrate of the protein chip;

[0035] The capture antibodies include: specific antibodies against interleukin-6, specific antibodies against tumor necrosis factor receptor II, specific antibodies against epiregulin and specific antibodies against interferon-inducible protein 10.

[0036] Preferably, immobilizing the capture antibody on the substrate of the protein chip comprises the following steps:

[0037] 100-1000pl of the solution of the captured antibody is spotted on the substrate, and after standing overnight at room temperature, it is vacuum-dried for 1-4 hours;

[0038] The solution of the capture antibody contains 0.02-20ng capture antibody and 0.01-10g / 100ml bovine albumin.

[0039] Among them, standing overnight at room temperature is conduc...

Embodiment 1

[0057] Example 1 Proteins associated with binding latent infection

[0058] Sex hormone-binding globulin (SHBG), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor receptor II ( TNF RII), epiregulin (Epiregulin), interferon-inducible protein 10 (IP-10), serine protease inhibitor (SERPINA3), tumor necrosis factor a (TNFa) protein standards, after gradient dilution, passed the prior art The fluorescent method was used to measure the fluorescence value, and the fluorescence signal value and corresponding concentration of the standard curve were obtained, as shown in Table 1.

[0059] Table 1 Fluorescence signal value and corresponding concentration of protein standard standard curve

[0060]

[0061]

[0062] From the data in the above table, logarithmically transform the concentration and fluorescence signal intensity respectively, draw a standard curve, and calculate a linear regression equation, wherein x is Log (concentration of sta...

Embodiment 2

[0080] Example 2 Preparation of a protein chip kit for the diagnosis of tuberculosis latent infection

[0081] In order to detect whether there are 4 kinds of proteins in the present invention in the sample, prepare the glass slide that is fixed with the specific antibody for following 4 kinds of proteins: interleukin-6 (IL-6), tumor necrosis factor receptor II (TNF RII), epiregulin (Epiregulin), interferon-inducible protein 10 (IP-10).

[0082] 1. The source of the antibody

[0083] Capture antibodies against specific proteins of interleukin-6 (IL-6), tumor necrosis factor receptor II (TNF RII), epiregulin (Epiregulin), interferon-inducible protein 10 (IP-10) The purpose, source and concentration of the detection antibody are detailed in Table 6. Among them, the capture antibody and the detection antibody recognize different epitopes of the protein, respectively.

[0084] The detection antibody is biotinylated, and the specific method is as follows: the detection antibody ...

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Abstract

The invention provides a kit and a protein chip for tuberculosis latent infection diagnosis. By detecting the expression levels of four proteins, i.e., interleukin-6, a tumor necrosis factor receptorII, an epidermal regulator and interferon-induced protein 10, tuberculosis latent infected people can be distinguished from healthy people, and the kit has high sensitivity and specificity. Specific antibodies corresponding to the four proteins are immobilized on a substrate to prepare the protein chip, the expression levels of the four proteins can be detected at the same time, and the protein chip has the advantages of low sample demand, simplicity and quickness. The application of the kit in molecular biological research and clinical detection of latent tuberculosis infection is beneficialto early diagnosis of latent tuberculosis infection, and latent tuberculosis infection patients can be found more accurately.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a kit and protein chip for the diagnosis of tuberculosis latent infection. Background technique [0002] Tuberculosis is the infectious disease with the largest number of deaths caused by a single pathogen after AIDS. One of the main reasons for the severe epidemic of tuberculosis is the lack of effective early diagnosis technology. [0003] After tuberculosis infection, not all people will get sick, and some people will turn into a state of latent infection, that is, latent tuberculosis infection. This part of the population continues to have tuberculosis in the body, but due to the control of the body's immunity to tuberculosis, there is no clinical manifestation. When the body's immunity is weakened, the tuberculosis bacteria lurking in the body will "resurrection", and the tuberculosis bacteria will replicate in the body, and finally turn into a tuberculosis patient. Th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/543G01N33/531
CPCG01N33/531G01N33/543G01N33/5695G01N33/6863G01N33/6869
Inventor 黄若磐杨倩婷张明霞陈骑张惠华易玉华
Owner RAYBIOTECH INC GUANGZHOU