Kit for rapidly detecting active tuberculosis

An active tuberculosis and kit technology, applied in the field of detection, can solve the problem of low specificity of the kit, and achieve the effect of wide coverage and practicability

Inactive Publication Date: 2020-01-17
GUANGZHOU DEAOU MEDICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specificity of the existing gamma-interferon release test kits is not high, only about 70%-85%, and there is still the possibility and demand for further improvement

Method used

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  • Kit for rapidly detecting active tuberculosis
  • Kit for rapidly detecting active tuberculosis
  • Kit for rapidly detecting active tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] (1) Sample collection:

[0043] In this example, a patient with active tuberculosis was taken as the experimental object, and no less than 15 mL of peripheral blood was collected in a heparin sodium blood collection tube, mixed upside down, and set aside;

[0044] (2) Lymphocyte separation: Use commercially available lymphocyte separation medium to separate lymphocytes in peripheral blood samples, take 4 mL of normal saline and mix them with 4 mL of peripheral blood, and slowly add them to the lymphocyte separation liquid. Centrifuge at a speed of 600r / min for 20min, absorb the cloud layer cells and make up to 12mL with 1640 medium (RPMI-1640 medium), centrifuge at a speed of 600r / min at room temperature for 10min, discard the supernatant, and make up the sediment to 5mL with 1640 medium And mix gently, centrifuge at 350r / min at room temperature for 10min, resuspend the cells with 1640 medium or PBS phosphate-buffered saline solution, then separate and purify the lympho...

Embodiment 2

[0052] (1) Sample collection:

[0053] In this example, a healthy person whose PPD test result was negative was taken as the experimental subject, and no less than 15 mL of peripheral blood was collected in a heparin sodium blood collection tube, mixed upside down, and set aside;

[0054] (2) Lymphocyte separation: Use commercially available lymphocyte separation medium to separate lymphocytes in peripheral blood samples, take 4 mL of normal saline and mix them with 4 mL of peripheral blood, and slowly add them to the lymphocyte separation liquid. Centrifuge at a speed of 600r / min for 20min, absorb the cloud layer cells and make up to 12mL with 1640 medium (RPMI-1640 medium), centrifuge at a speed of 600r / min at room temperature for 10min, discard the supernatant, and make up the sediment to 5mL with 1640 medium And mix gently, centrifuge at 350r / min at room temperature for 10min, resuspend the cells with 1640 medium or PBS phosphate-buffered saline solution, then separate and...

Embodiment 3

[0063] (1) Sample collection:

[0064] In this embodiment, 10 cases of active tuberculosis patients, 10 cases of tuberculosis infection (PPD test results are positive) and 10 healthy people (PPD test results are negative), a total of 30 samples are used as experimental subjects. Peripheral blood less than 12mL, mix upside down and set aside;

[0065] (2) Lymphocyte separation: Use commercially available lymphocyte separation medium to separate lymphocytes in peripheral blood samples, take 4 mL of normal saline and mix them with 4 mL of peripheral blood, and slowly add them to the lymphocyte separation liquid. Centrifuge at a speed of 600r / min for 20min, absorb the cloud layer cells and make up to 12mL with 1640 medium (RPMI-1640 medium), centrifuge at a speed of 600r / min at room temperature for 10min, discard the supernatant, and make up the sediment to 5mL with 1640 medium And mix gently, centrifuge at 350r / min at room temperature for 10min, resuspend the cells with 1640 med...

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Abstract

The invention belongs to the field of detection, and particularly relates to a kit for rapidly detecting active tuberculosis. The kit comprises a stimulating protein and a stimulant, a mixed stimuluscomposed of the stimulating protein and the stimulant is used as a specific stimulating antigen to stimulate peripheral blood lymphocytes to produce cell factors IFN- gamma and IL-2, and the purpose of judging whether the disease is the active tuberculosis is achieved by detecting the secretion concentration of IFN- gamma and IL-2. The stimulating protein of the kit provided by the invention comprises CFP-10, ESAT-6 and Rv 1985c, and the stimulant comprises a phorbol ester polyclonal stimulant and ionomycin. The stimulating protein can stimulate tuberculosis-specific effector T cells in the lymphocytes to secrete the cell factors, and the stimulant can stimulate the activated lymphocytes to enhance the ability of synthesizing and secreting the cell factors. By adopting the kit provided bythe invention, the specificity of the detection result can be greatly improved, and whether the disease is the active tuberculosis can be judged more accurately.

Description

technical field [0001] The invention belongs to the detection field, in particular to a kit for rapidly detecting active tuberculosis. Background technique [0002] Tuberculosis is a common chronic infectious disease caused by Mycobacterium tuberculosis, which can invade many organs, with pulmonary tuberculosis infection being the most common. At present, nearly 1 / 3 of the world's people are infected with Mycobacterium tuberculosis. Statistics show that in 2013, 1.5 million people died of tuberculosis, and 9 million new cases occurred. Tuberculosis has become one of the major diseases that cause adult deaths from infectious diseases all over the world. China is one of the 22 countries with a high burden of tuberculosis in the world, and the number of active tuberculosis patients ranks second in the world. The transmission of tuberculosis mainly occurs before the patient is discovered and treated. One tuberculosis patient can infect as many as 10-15 people through close cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/6866G01N33/6869G01N33/5695G01N2333/35G01N2333/55G01N2333/57
Inventor 胡鹏南葛泰宏何鑫肖艳文
Owner GUANGZHOU DEAOU MEDICAL TECH CO LTD
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