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
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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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