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Method for detecting pathogenic microorganism by using antigen-stimulated cellular immune response and test pen

A technology of pathogenic microorganisms and cellular immunity, applied in the fields of immunology, protein detection and molecular biology, can solve the problems of delayed treatment time, low content of pathogenic microorganisms, complicated process, etc., to improve specificity and sensitivity, reduce manpower and material resources, simple method effect

Inactive Publication Date: 2011-04-27
ABBOTT DIAGNOSTICS (SHANGHAI) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most commonly used methods are to detect antibodies, antigens, DNA or RNA of pathogenic microorganisms in body fluids, but it is difficult to detect pathogenic microorganisms due to the small amount of pathogenic microorganisms in the initial stage, and it takes a long time and the process is complicated to take samples and culture and amplify microorganisms , not only is easy to introduce human error but also often delays the best treatment time, so it is necessary to develop a fast, simple and effective detection method to detect diseases caused by such pathogenic microorganisms

Method used

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  • Method for detecting pathogenic microorganism by using antigen-stimulated cellular immune response and test pen
  • Method for detecting pathogenic microorganism by using antigen-stimulated cellular immune response and test pen
  • Method for detecting pathogenic microorganism by using antigen-stimulated cellular immune response and test pen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, the preparation of the colloidal gold immunochromatography test pen that detects IFN-γ

[0030] figure 1 The preparation method of the test pen shown is as follows:

[0031] 1. Preparation of colloidal gold: Trisodium citrate reduction method (HAuCl 4 , Trisodium citrate was purchased from Sigma Company) to prepare colloidal gold, and the particle size of colloidal gold was about 40nm.

[0032] 2. Preparation of Anti-IFN-γ Antibody Colloidal Gold Pads

[0033] Use colloidal gold to label anti-IFN-γ monoclonal antibody (Abcam) at a ratio of 1mg antibody / 1500D colloidal gold at pH 8.4. After 30 minutes, add an appropriate amount of BSA at pH 7.4 and concentrate by centrifugation After that, the IFN-γ antibody gold probe was obtained.

[0034] The gold probe was diluted to OD2.0 with colloidal gold film diluent (0.1M TAPS buffer, 20% sucrose, 3.75% BSA, pH 8.0) for use.

[0035] After fully immersing the glass fiber (Millipore company) in the IFN-γ collo...

Embodiment 2

[0046] Example 2, detect Mycobacterium tuberculosis by detecting IFN-γ in human blood

[0047] In the first step, a blood sample is collected. Take 1ml of blood from the patient and add it to a sterile test tube containing heparin (Hong Kong Advanced Technology Industries Co., Ltd.) as a negative control tube; ) in a sterile test tube as a measuring tube; then draw 1ml of blood from the patient, and add heparin and cell mitogens (such as phytohemagglutinin, Hong Kong Advanced Technology Industries Co., Ltd.) into a positive control tube. Shake the 3 tubes of blood fully respectively. Start the detection test within 12 hours. The second step is to incubate the sample. After blood samples were collected, 3 tubes were incubated at 37°C for 6-24 hours within 12 hours. The third step is sample testing. Take 100 μl sample dropwise into figure 1 The colloidal gold immunochromatographic test pen for detecting IFN-γ (included in the box) figure 2 If there is a red line at the c...

Embodiment 3

[0052] Embodiment 3, detect HIV by detecting IFN-γ in human blood

[0053] In the first step, a blood sample is collected. Take 1ml of blood from the patient and add it to a sterile test tube containing heparin (Hong Kong Advanced Technology Industries Co., Ltd.) as a negative control tube; Industrial Co., Ltd.) as a measuring tube; then draw 1ml of blood from the patient, and add heparin and mitogens (such as phytohemagglutinin, phytohemagglutinin, Hong Kong Advanced Technology Industries Co., Ltd.) as a positive control tube. Shake the 3 tubes of blood fully respectively. Start the detection test within 12 hours. The second step is to incubate the sample. After blood samples were collected, 3 tubes were incubated at 37°C for 6-24 hours within 12 hours. The third step is sample testing. Take 100 μl sample dropwise into figure 1 The test pen for detecting IFN-γ as shown (included in the box) figure 2 Show the colloidal gold immunochromatography test strip that detects ...

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Abstract

The invention discloses a method for detecting a pathogenic microorganism by using antigen-stimulated cellular immune response. In the method, immune cells, which have been exposed to the microorganism, in blood are stimulated by specific antigen of the pathogenic microorganism or a functional segment of the pathogenic microorganism, so a cell factor, such as interferon gamma (IFN-gamma), is produced by the immune cells; and then whether the pathogenic microorganism exists in animals is judged by detecting the amount of the IFN-gamma. In the invention, the IFN-gamma in the blood is detected by a colloidal gold-labeled immunochromatographic assay. In addition, the invention also discloses a colloidal gold immunochromatographic assay test pen for detecting the pathogenic microorganism. By the method, the pathogenic microorganism in an incubation period or early morbidity can be detected quickly and effectively.

Description

technical field [0001] The invention relates to the technical fields of molecular biology, immunology and protein detection. More specifically, it relates to a method for detecting pathogenic microorganisms by using antigen-stimulated cellular immune response; in addition, the present invention also relates to a colloidal gold immunochromatographic test pen for detecting pathogenic microorganisms. Background technique [0002] There are many diseases caused by pathogenic microorganism infection, such as tuberculosis, AIDS (AIDS), bird flu, severe acute respiratory syndrome (severe acute respiratory syndrome, SARS), hepatitis, malaria, anthrax, pneumonia, hepatitis, mad cow disease, etc., these diseases Because it can seriously endanger personal health and is highly contagious, it poses a great threat to human health. In addition, because these pathogenic microorganisms can hide in the body for a long time in many cases without onset or mild onset, it has brought great obsta...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/577G01N21/76
CPCG01N33/54386G01N2333/195G01N2333/05G01N33/54388
Inventor 杨吉杨挥
Owner ABBOTT DIAGNOSTICS (SHANGHAI) CO LTD
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