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Kit for detecting mycobacterium tuberculosis infection by using peripheral blood and application of kit

A technology of Mycobacterium tuberculosis and a kit, which is applied in measurement devices, analytical materials, biological material analysis, etc., can solve the problems that diagnostic products are not suitable for clinical diagnostic applications, high cost, etc., and achieve high clinical application value, easy operation, high cost, etc. Effects with high sensitivity and specificity

Active Publication Date: 2014-04-30
北京同生时代生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, due to China's special tuberculosis epidemiology and ecological environment, and the high cost of patent protection, these diagnostic products are not suitable for a wide range of clinical diagnostic applications in China. Therefore, it is urgent to develop tuberculosis rapid diagnostic reagents with independent intellectual property rights

Method used

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  • Kit for detecting mycobacterium tuberculosis infection by using peripheral blood and application of kit
  • Kit for detecting mycobacterium tuberculosis infection by using peripheral blood and application of kit
  • Kit for detecting mycobacterium tuberculosis infection by using peripheral blood and application of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0061] Example 1. Development of a kit for detecting Mycobacterium tuberculosis infection using peripheral blood

[0062] The kit provided by the present invention utilizes peripheral blood to detect Mycobacterium tuberculosis infection and comprises:

[0063] 1. Mycobacterium tuberculosis Rv3615c mixed peptide library;

[0064] The Mycobacterium tuberculosis Rv3615c mixed polypeptide peptide library consists of 19 polypeptides consisting of amino acid sequences shown in sequences 1-19 in the sequence listing;

[0065] 2. Positive control stimulus-phytohemagglutinin (PHA) solution;

[0066] 3. 24-well plate for cell culture;

[0067] 4. FITC (fluorescein isothiocyanate) labeled human CD3 monoclonal antibody, R-PE (R-phycoerythrin, R-phycoerythrin) labeled human γ-interferon monoclonal antibody;

[0068] 5. Golgi blocker (BrefeldinA);

[0069] 6. Cell fixative and permeabilization buffer.

[0070] 7. Other reagents and consumables required for flow detection.

Embodiment 2

[0071] Embodiment 2, the kit of application embodiment 1 carries out clinical detection of Mycobacterium tuberculosis infection

[0072] 1. Collection of peripheral blood samples

[0073] The screening criteria for volunteer cases are to meet the following clinical diagnostic criteria at the same time:

[0074] (1) The clinical attending doctor is diagnosed with pulmonary tuberculosis according to the clinical manifestations of the volunteers;

[0075] (2) Mycobacterium tuberculosis isolated and cultured colonies are positive;

[0076] (3) X-ray images showing lung lesions;

[0077] (4) The result of tuberculin subcutaneous test is positive, and the spot diameter is greater than 1cm.

[0078] Healthy volunteers are screened and marked as meeting the following at the same time:

[0079] (1) No clinical symptoms of tuberculosis;

[0080] (2) No other diseases or infections.

[0081] After the selected individuals pass the physical examination of the clinician, the experime...

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Abstract

The invention discloses a kit for detecting mycobacterium tuberculosis infection by using peripheral blood. The kit provided by the invention comprises the following substances: (1) a mycobacterium tuberculosis Rv3615c mixed polypeptide library which consists of all or a part of 19 polypeptides consisting of amino acid sequences shown as sequences 1 to 19 in a sequence table, (2) an anti-CD3 antibody labeled with a fluorescent dye A, (3) an anti-gamma-interferon antibody labeled with a fluorescent dye B, and (4) a Golgi apparatus blocking agent, wherein the fluorescent dye A and the fluorescent dye B emit different fluorescent colors. A novel method which can be used for detecting mycobacterium tuberculosis infection is provided. As proved by experiments, antigenic stimulation can be performed by directly using the peripheral blood without separating single karyocytes of the peripheral blood. As indicated by experimental data, the kit has high sensitivity and high specificity when being used for detecting mycobacterium tuberculosis infection. Moreover, the kit is easy and convenient to operate and low in cost, and has high clinical application value.

Description

technical field [0001] The invention relates to a kit for detecting mycobacterium tuberculosis infection by using peripheral blood and its application, in particular to a kit for detecting mycobacterium tuberculosis infection by using peripheral blood based on flow cytometry and its application. Background technique [0002] Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis. According to WHO estimates, about one-third of the world's population is subtly infected by Mycobacterium tuberculosis. There are more than nine million new cases of active infection and one million deaths each year. example above. In recent years, the incidence of tuberculosis in my country has remained high, with more than one million new cases and nearly 200,000 deaths every year, which has brought serious harm to the health of our citizens, social stability and economic development. [0003] Early diagnosis of tuberculosis is of great significance in controlling the disease ...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/5695G01N33/6884G01N33/6893G01N2800/12
Inventor 石佳李启靖董剑峰张卫红
Owner 北京同生时代生物技术有限公司
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