Blood miR-3135b detection kit used for diagnosis of coronary artery vascular calcification
A technology for blood detection and vascular calcification, applied in the field of blood detection kits and diagnostic blood detection kits
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Embodiment 1
[0044] A significant positive correlation was confirmed between the expression of miRNA-3135b in serum and the integral level of coronary artery calcium detected in patients.
[0045] Preparation of plasma samples
[0046] 1). Use EDTA anticoagulant tubes to collect blood from the subject, store it at room temperature or at 4 degrees, and proceed to the next step within one hour.
[0047] 2). Centrifuge at 1900g (3000rpm) at 4 degrees for 10 minutes
[0048] 3). Take the upper yellow liquid as plasma
[0049] 4). Centrifuge at 16000g for 10 minutes
[0050] 5). Take the supernatant and store it at -80°C for later use.
[0051] Extraction of miRNA from plasma
[0052] Use miRNeasy Serum / Plasma kit (217184) and miRNeasy Serum / Plasma Spike-in-control (219610) from QIAGEN, USA.
[0053] The specific method is as follows:
[0054] 1). Thaw the serum stored at -80 degrees, and take 200ul each.
[0055] 2). Add 1ml of QIAzol Lysis Reagent (provided with the kit), and mix thoro...
Embodiment 2
[0089] Example 2 Detection rate verification
[0090] Subject recruitment criteria:
[0091] Male aged 50-70 years old, in good health, 162-175cm in height, no drug dependence, no mental, psychological disease, genetic disease and infectious disease history; no history of severe allergies; no malnutrition.
[0092] Clinical detection indicators:
[0093] (1) Coronary CT angiography, calculate coronary artery calcification score
[0094] (2) Detection of expression level of miRNA-3135b in serum
[0095] The extraction, detection and result analysis of miR-3135b in the subject's serum are the same as in Example 1.
[0096] Experimental results:
[0097] According to the current clinical coronary artery calcification detection standard, through coronary CT detection, a CAS greater than 100 is a patient with coronary artery calcification. In this study, the serum of 200 subjects was collected and tested according to the above-mentioned Example 1. The microRNA copy number was ...
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