Kit for detecting juvenile osteoporosis

Abstract

The invention discloses a kit for detecting juvenile osteoporosis. According to the kit, an obvious mutant site which is a site 1020 is discovered to exist in a DKK1 gene in a patient suffering from the juvenile osteoporosis and a normal crowd through genetic comparison. Through detecting the mutant site, particularly provided one pair of specific primer pairs, a human body which suffers from the juvenile osteoporosis can be specifically identified. Compared with the method in the prior art, a detection method has the advantages of being quick for detection, accurate and quick, and is suitable to popularize and use.

Description

technical field

[0001] The invention belongs to the detection field and relates to a kit for juvenile osteoporosis detection. Background technique

[0002] Osteoporosis (osteoporosis) is a systemic bone disease in which bone density and bone quality decrease due to various reasons, bone microstructure is destroyed, bone fragility increases, and fractures are prone to occur. In the early years, it was generally believed that osteopenia was osteoporosis, while in the United States, fractures in the elderly were considered osteoporosis. It was not until the Third International Osteoporosis Symposium held in Denmark in 1990 and the Fourth International Osteoporosis Symposium held in Hong Kong in 1993 that osteoporosis had a clear definition and was recognized worldwide. Background: Primary osteoporosis is a systemic skeletal disease characterized by decreased bone mass and degeneration of bone microstructure, resulting in increased bone fragility and prone to fractures. Every ...

Examples

Embodiment 1

[0026] Embodiment one: see figure 1 As shown, Example 1 screening of juvenile osteoporosis patients and screening of gene variation in normal people:

[0027] 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:

[0028] 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, or pulmonary insufficiency, no various endocrine diseases causing secondary osteoporosis, no tumor, diabetes and other serious diseases that interfere with bone metabolism.

[0029] Normal group: select 20 healthy volunteers aged 8-15, 10 males and 10 males.

[0030] There was no significan...

Embodiment 2

[0034] Embodiment two: PCR amplification identification

[0035] (1) Primer design

[0036]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.

[0037] 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;

[0038] PCR reaction conditions: pre-denaturation at 95°C for 5 min, entering a cycle of 95°C for 60 s, 57.5°C for 1 min and 72°C for 1 min, 35 cycles, and finally extending at 72°C for 5 min.

[0039] The polymerase chain reaction system was purchased from Shanghai Sangong.

[0040] (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 u...

technical field

[0001] The invention belongs to the detection field and relates to a kit for juvenile osteoporosis detection. Background technique

[0002] Osteoporosis (osteoporosis) is a systemic bone disease in which bone density and bone quality decrease due to various reasons, bone microstructure is destroyed, bone fragility increases, and fractures are prone to occur. In the early years, it was generally believed that osteopenia was osteoporosis, while in the United States, fractures in the elderly were considered osteoporosis. It was not until the Third International Osteoporosis Symposium held in Denmark in 1990 and the Fourth International Osteoporosis Symposium held in Hong Kong in 1993 that osteoporosis had a clear definition and was recognized worldwide. Background: Primary osteoporosis is a systemic skeletal disease characterized by decreased bone mass and degeneration of bone microstructure, resulting in increased bone fragility and prone to fractures. Every ...

Examples

Embodiment 1

[0026] Embodiment one: see figure 1 As shown, Example 1 screening of juvenile osteoporosis patients and screening of gene variation in normal people:

[0027] 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:

[0028] 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, or pulmonary insufficiency, no various endocrine diseases causing secondary osteoporosis, no tumor, diabetes and other serious diseases that interfere with bone metabolism.

[0029] Normal group: select 20 healthy volunteers aged 8-15, 10 males and 10 males.

[0030] There was no significan...

Embodiment 2

[0034] Embodiment two: PCR amplification identification

[0035] (1) Primer design

[0036]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.

[0037] 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;

[0038] PCR reaction conditions: pre-denaturation at 95°C for 5 min, entering a cycle of 95°C for 60 s, 57.5°C for 1 min and 72°C for 1 min, 35 cycles, and finally extending at 72°C for 5 min.

[0039] The polymerase chain reaction system was purchased from Shanghai Sangong.

[0040] (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 u...

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