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A protein chip and kit for active tuberculosis diagnosis

A technology of active tuberculosis and protein chips, applied in biological testing, instruments, measuring devices, etc., to achieve the effect of less specimen consumption, high sensitivity and specificity, and increased stability

Active Publication Date: 2019-01-04
深圳市星系生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the use of protein chips to screen tuberculosis-specific molecular markers for diagnosis or treatment.

Method used

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  • A protein chip and kit for active tuberculosis diagnosis
  • A protein chip and kit for active tuberculosis diagnosis
  • A protein chip and kit for active tuberculosis diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Determination of protein chip spotting conditions

[0023] 1. Spotting concentration of the capture antibody: the capture antibody (specific antibody for 29 kinds of proteins) in different buffers (sterile water, PBS buffer, PBS containing different concentrations of bovine albumin, PBS containing different concentrations of glycerol) PBS buffer solution, etc.) was diluted at a concentration of 2x, spotted on the surface of the chip with a non-contact spotter, and then compared its activity and stability with a sandwich ELISA method. Experiments show that diluting the capture antibody with PBS buffer containing 0.01-10g / 100ml bovine albumin has the best linear rate and stability.

[0024] 2. Condition control of spotting: when the capture antibody (specific antibody for 29 kinds of proteins) is directly spotted on the surface of the glass slide at room temperature, the spot of the capture antibody is prone to produce hollow spots, and the activity of differen...

Embodiment 2

[0025] Embodiment 2: the preparation of the protein chip kit that is used for active tuberculosis diagnosis of the present invention.

[0026] In order to detect whether there are 29 kinds of proteins in the present invention in the sample, prepare the glass slides that are fixed with specific antibodies against the following 29 kinds of proteins: integrin-like metalloprotease and thrombin 8 (ADAM8), carbohydrate sulfotransferase 4 (CHST4), cytotoxic T lymphocyte-associated antigen 4

[0027](CTLA4), cysteine ​​protease inhibitor E / M (Cystatin E / M), folate receptor 2 (FOLR2), granulocyte colony-stimulating biological factor (GM-CSF), hyaluronan proteoglycan connexin 1 (HAPLN1), interferon alpha / beta receptor 2 (IFNab R2), insulin-like growth factor binding protein 5 (IGFBP-5), interleukin-1R6 (IL-1R6), interleukin-11 (IL-11 ), Interleukin-12p40 (IL-12p40), Interleukin-21R (IL-21R), Interleukin-23 (IL-23), Interleukin-29 (IL-29), Interleukin- 7 (IL-7), kallikrein 14 (Kallikre...

Embodiment 3

[0044] Embodiment 3: the preparation of the polypeptide mixture that stimulates peripheral blood

[0045] In order to ensure the specificity of the stimulating polypeptide, the sequences of two polypeptides were designed according to the CFP-10 and ESAT-6 proteins, see Table 3 for details. The peptide synthesis is completed by Shenzhen Hanyu Biotechnology Co., Ltd., and the purity of the peptide exceeds 95%.

[0046] The source of table 3 polypeptide and and sequence list

[0047]

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PUM

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Abstract

The invention discloses a protein chip and a kit for abortive tuberculosis diagnosis. The protein chip contains a specific antibody for 29 proteins. The protein chip can specifically separate an abortive tuberculosis patient from healthy people, nontuberculous pneumonia patients and the healed tuberculosis patients, has relatively high sensitivity and specificity, high flux and less consumption of specimens and is convenient in popularization and mass production in ordinary laboratories; and meanwhile, the application in the clinical diagnosis is favorable for more rapidly and accurately discovering the abortive tuberculosis patient and is expected to be used for screening the abortive tuberculosis and diagnosing the abortive tuberculosis, inspecting the healthy people, predicting and evaluating the curative effect of the tuberculosis treatment and the like, and provides strong technical support for controlling the situation of the tuberculosis.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a protein chip kit for active tuberculosis diagnosis. Background technique [0002] According to the World Health Organization data in 2015, tuberculosis is the infectious disease with the largest number of deaths caused by a single pathogen after AIDS. How to reduce the tuberculosis epidemic is a major issue facing the economic and social development of our city. One of the main reasons for the severity of the tuberculosis epidemic is the lack of effective early diagnosis techniques. [0003] In terms of tuberculosis diagnostic technology, more than a century has passed, and we still use the earliest tuberculosis diagnostic technology in clinical practice today: tuberculosis smear and tuberculosis culture. Although the operation is simple and the price is economical, the sensitivity of smear does not exceed 30%; the sensitivity of culture does not exceed 40%, and because i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/68
CPCG01N33/56911G01N33/68
Inventor 杨帆刘小山黄若磐
Owner 深圳市星系生物科技有限公司
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