Application of carbonic anhydrase-2 as a detection marker in the diagnosis of kidney stones
A carbonic anhydrase, detection label technology, applied in the field of biomedical diagnosis
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Embodiment 1
[0036] Select 3 patients with kidney stones and 3 normal patients, and collect the urine of the above patients for the screening of proteomics technology. The above-mentioned people are all male, and their age and weight are roughly similar.
[0037] The inventor screened 8 proteinuria markers in the early stage, of which only carbonic anhydrase-2 was stably expressed in the urine of patients with kidney stones.
[0038] Inclusion criteria of the test population: the inventor included healthy people, patients with kidney stones and patients with kidney tumors from January 2017 to October 2018. The following implementation steps can only be carried out after the above-mentioned population meets the following criteria:
[0039] (1) Age 18-68;
[0040] (2) Urinary tract color Doppler ultrasound, X-ray and CT examinations were performed to confirm the diagnosis of patients with kidney stones;
[0041] (3) All patients with renal tumors underwent surgery and were diagnosed throug...
Embodiment 2
[0056] The detection method of carbonic anhydrase-2 content and activity, comprises the following steps:
[0057] (1) Collect the early morning urine of the subject and send it for inspection within 2 hours. If it cannot be sent for inspection in time, it should be stored at 5°C, but the storage time is required to be less than 24 hours. The urine needs to be centrifuged at 1800r / min before testing. After 10 minutes, take supernatant samples for testing;
[0058] (2) Take out the microwell ELISA plate, set standard wells and sample wells, add 50 μL of carbonic anhydrase-2 standard substances of different concentrations to each of the standard wells, and add 50 μL of the sample to be tested in the sample wells;
[0059] (3) Add 100 μL of horseradish peroxidase-labeled carbonic anhydrase-2 antibody to each well of standard wells and sample wells, and incubate in a 37°C water bath for 65 minutes;
[0060] (4) Discard the liquid, pat dry on absorbent paper, fill each well with wa...
Embodiment 3
[0065] The detection method of carbonic anhydrase-2 content and activity, comprises the following steps:
[0066] (1) Collect the early morning urine of the subject and send it for inspection within 2 hours. If it cannot be sent for inspection in time, it should be stored at 6°C, but the storage time is required to be less than 24 hours. The urine needs to be centrifuged at 2000r / min before testing. 6min, take the supernatant sample to be detected;
[0067] (2) Take out the microwell ELISA plate, set standard wells and sample wells, add 50 μL of carbonic anhydrase-2 standard substances of different concentrations to each of the standard wells, and add 50 μL of the sample to be tested in the sample wells;
[0068] (3) Add 100 μL of horseradish peroxidase-labeled carbonic anhydrase-2 antibody to each well of standard wells and sample wells, and incubate in a 37°C water bath for 65 minutes;
[0069] (4) Discard the liquid, pat dry on absorbent paper, fill each well with washing ...
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