Kit for juvenile osteoporosis detection
A technology of osteoporosis and kit, which is applied in the field of detection, can solve the problems that cannot be popularized and used in a large area, and achieve the effect of convenient, timely and accurate genetic diagnosis and detection, and reduce the deformity rate
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Embodiment 1
[0027] Embodiment one: see figure 1 , 2 As shown, Example 1 screening of juvenile osteoporosis patients and screening of gene variation in normal people:
[0028] 1. Research objects: Case samples were collected from the First Affiliated Hospital of Soochow University and the Second Affiliated Hospital of Soochow University, as follows:
[0029] Juvenile osteoporosis group: 150 juvenile osteoporosis patients admitted to the orthopedics department of the hospital were selected, 25 males and 25 females, the youngest was 20 years old and the oldest was 50 years old. Inclusion criteria: in line with the "Recommended Diagnostic Criteria for Osteoporosis in China". No obvious heart, liver, kidney, pulmonary insufficiency, no various endocrine diseases causing secondary osteoporosis, no tumor, diabetes and other serious diseases that interfere with bone metabolism.
[0030] Normal group: select 20 healthy volunteers aged 8-15, 10 males and 10 males.
[0031] There was no signific...
Embodiment 2
[0035] Embodiment two: PCR amplification identification
[0036] (1) Primer design
[0037] PCR amplification primers were designed according to the sequence of DKK-1 gene AF177394.1 in Genbank, and synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. The specific primer sequences are shown in SEQ ID NO: 1-2. Housekeeping genes were used as controls.
[0038] The PCR reaction system is: DNA template 1 μl, upstream and downstream primers 0.5 μl each, PCR Mix 12.5 μl, ultrapure water supplemented to 25 μl;
[0039] PCR reaction conditions: pre-denaturation at 95°C for 5min, entering a cycle of 95°C for 60s, 57.5°C for 1min and 72°C for 1min, 35 cycles, and finally extending at 72°C for 5min.
[0040] The polymerase chain reaction system was purchased from Shanghai Sangong.
[0041] (2) Acquisition of templates: the blood DNA of another 30 groups of juvenile osteoporosis patients and 10 groups of normal people was extracted, and the specific method used a ...
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