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Application of CFAP20 gene in diagnosis of osteoporosis

A technology for osteoporosis and genes, which is applied in the detection/testing of microorganisms, chemical libraries, combinatorial chemistry, etc., can solve the problems that patients with osteoporosis cannot be screened, and achieve early diagnosis, genetic diagnosis, and more The timely effect of genetic diagnosis

Inactive Publication Date: 2017-08-15
TAISHAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The objects of auxiliary examination are generally patients with advanced osteoporosis, and cannot be used for the screening of patients with early osteoporosis

Method used

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  • Application of CFAP20 gene in diagnosis of osteoporosis
  • Application of CFAP20 gene in diagnosis of osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening of genes differentially expressed in patients with osteoporosis and normal people

[0054] 1. Research object:

[0055] Osteoporosis group: Randomly select 10 patients with primary osteoporosis from the orthopedics department of the hospital, aged 50-75 years.

[0056] Inclusion criteria: in line with the "Recommended Diagnostic Criteria for Osteoporosis in China" (Second Draft). 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.

[0057] Normal group: 10 healthy volunteers aged 50-75 were selected.

[0058] There was no significant difference in age and gender between the two groups (P>0.10), which were comparable.

[0059] All research subjects were informed about this study and signed the informed consent.

[0060] 2. Extraction of total RNA in blood

[0061] Total RNA in blood leukocytes was ...

Embodiment 2

[0075] Example 2 QPCR experiment verification of genes differentially expressed in patients with osteoporosis and normal people

[0076] 1. Research object:

[0077] According to the method of Example 1, 50 cases of osteoporosis patients and 50 normal persons were collected.

[0078] 2. Extraction of total RNA in blood

[0079] Method is with embodiment 1.

[0080] 3. Reverse transcription

[0081] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC water, 5× reverse transcription buffer, 10mmol / L dNTP, 0.1mmol / l DTT, 30μmmol / l Oligo dT, 200 U / μl M-MLV, template RNA. Incubate at 42°C for 1 hour, then centrifuge briefly at 72°C for 10 minutes.

[0082] 4. QPCR

[0083] (1) Primer design

[0084] QPCR amplification primers were designed according to the coding sequences of CFAP20 gene an...

Embodiment 3

[0100] The preparation of embodiment 3 osteoporosis diagnostic kits

[0101] According to the correlation between the CFAP20 gene and osteoporosis, the osteoporosis can be diagnosed by detecting the expression of the CFAP20 gene. Accordingly, the present invention provides a kit for diagnosing osteoporosis based on detecting the expression of the CFAP20 gene. The components in the diagnostic kit are as follows: a SYBR Green polymerase chain reaction system; a pair of primers for amplifying the CFAP20 gene and the GAPDH gene. The forward primer sequence for amplifying CFAP20 gene is 5'-ATACGGCACCAATTACAT-3', the reverse primer sequence is 5'-TAGAGTCTGTCTGAGAAGTA-3'; the forward primer sequence for amplifying GAPDH is 5'-TTTAACTCTGGTAAAGTGGATAT-3', reverse The primer sequence was 5'-GGTGGAATCATATTGGAACA-3'. SYBRGreen polymerase chain reaction system contains PCR buffer, dNTPs, SYBR Green fluorescent dye. The PCR buffer composition is: 25mM KCl, 2.5mMMgCl 2 , 200mM (NH 4 ) 2...

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Abstract

The invention discloses an application of a CFAP20 gene as a molecular marker in early diagnosis of osteoporosis. The invention researches the differential expression genes in patients with osteoporosis by using high flux sequencing and QPCR experiments. The research findings of the invention show that whether a subject suffers from osteoporosis can be judged through detecting the condition of CFAP20 gene expression in blood, and the detection method can be used for early diagnosis of osteoporosis and has the characteristics of timeliness and accuracy.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis, and relates to a molecular marker for diagnosing osteoporosis, in particular to the application of the molecular marker-CFAP20 gene in blood in the preparation of products for diagnosing osteoporosis. Background technique [0002] Osteoporosis is a common metabolic bone disease characterized by decreased bone mass, destruction of bone microarchitecture, increased bone fragility, and susceptibility to fracture. It is caused by excessive bone resorption due to increased / over-activated osteoclasts, or decreased bone formation caused by osteoblast reduction / functional defects, resulting in excessive bone loss, and is one of the most important diseases that harm middle-aged and elderly people . [0003] The detection of osteoporosis includes the detection of laboratory examination indicators and auxiliary detection. [0004] Laboratory inspection indicators: [0005] Some serum biochemical indica...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06C40B40/10G01N33/53
CPCC12Q1/6841C12Q1/686C12Q1/6869C12Q1/6883C12Q2600/158C40B40/06C40B40/10G01N33/53C12Q2561/113C12Q2545/114C12Q2521/107C12Q2535/122
Inventor 娄志国苗芳
Owner TAISHAN MEDICAL UNIV
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