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Molecular marker relevant to adolescent idiopathic scoliosis and application thereof

A technology of molecular markers and scoliosis, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve problems such as inconsistent conclusions, inability to occur AIS, and satisfactory interpretation of progress

Inactive Publication Date: 2018-04-06
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a large number of researches on its genomics and proteomics at home and abroad have achieved certain results, the conclusions are not consistent, and it is impossible to make a satisfactory explanation for the occurrence and progress of AIS.

Method used

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  • Molecular marker relevant to adolescent idiopathic scoliosis and application thereof
  • Molecular marker relevant to adolescent idiopathic scoliosis and application thereof
  • Molecular marker relevant to adolescent idiopathic scoliosis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Sample collection standard

[0038] 1. Research object

[0039] This study adopts a case-control research method. The case group of peripheral blood samples is selected from Chinese Han female AIS who underwent scoliosis correction in Peking Union Medical College Hospital.

[0040] Paraspinal muscle specimens The case group was selected from Chinese Han female AIS patients who underwent scoliosis correction in Peking Union Medical College Hospital, and the control group was the intervertebral disc degeneration surgery patients without spinal deformity matched with the case group in terms of gender, age, and ethnicity.

[0041] 1.1 Case group

[0042] All come from adolescent idiopathic scoliosis patients undergoing surgical treatment in our hospital from April 2013 to March 2014. Complete clinical data and imaging data (including spinal X-ray, CT and three-dimensional reconstruction, total spine MRI) were selected. ) Han adolescent idiopathic scoliosis patie...

Embodiment 2

[0066] Example 2 Screening of lncRNA

[0067] Four cases of adolescent idiopathic scoliosis patients were taken as the AIS group, and four normal controls were taken as the normal control group (NC group). Fasting peripheral blood was collected and total RNA was extracted.

[0068] 1. Extraction of total RNA from peripheral whole blood

[0069] 1) Add 0.25mL peripheral blood and 0.75mL Trizol LS reagent cryopreservation mixture to each EP tube, shake vigorously to mix;

[0070] 2) Incubate at room temperature for 5 minutes to completely dissociate the nucleic acid-protein complex;

[0071] 3) Add 0.2mL chloroform to each EP tube (that is, add 0.2mL chloroform to every 0.75mL Trizol LS reagent reagent sample), and cap the EP tube tightly. Shake vigorously for 15 seconds;

[0072] 4) Stand at room temperature for 3 minutes, so that the organic phase and the inorganic phase can be separated quickly and effectively;

[0073] 5) Centrifuge at 4°C and 12000G for 10 minutes in a ...

Embodiment 3

[0152] Example 3 qRT-PCR large sample verification

[0153] Taking 20 age-matched AIS patients and 20 normal females as the research objects, ENST00000602322.1 and MIR210HG with obvious differential expression and predicted to be involved in the pathogenesis of AIS were selected for real-time quantitative PCR verification.

[0154] 1. Extraction of total RNA from peripheral whole blood

[0155] The steps are shown in Example 2.

[0156] 2. The reverse transcription process of cDNA synthesis:

[0157] Reverse transcribe the qualified RNA into cDNA, the specific steps are as follows:

[0158] 1) Removal of DNA in the RNA sample: add the reagents in Table 8 below to the centrifuge tube in turn, and vortex to mix.

[0159] 2) Slightly centrifuge, place in a PCR instrument, and react at 42°C for 5 minutes;

[0160] 3) Prepare the reverse transcription system as shown in Table 9, add it to the centrifuge tube that has completed step 2, and vortex to mix;

[0161] 4) Slightly ce...

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Abstract

The invention discloses a molecular marker ENST00000602322.1 and / or MIR210HG relevant to adolescent idiopathic scoliosis, and further verifies that ENST00000602322.1 represents down-regulated repression in adolescent idiopathic scoliosis, and MIR210HG represents up-regulated expression in adolescent idiopathic scoliosis, and therefore, the molecular marker can be applied to preparation of productsfor diagnosing or predicating adolescent idiopathic scoliosis. Meanwhile, the invention further discloses an application of the molecular marker in preparation of a kit for diagnosing or predicatingadolescent idiopathic scoliosis. By using the molecular marker, early diagnosis of adolescent idiopathic scoliosis can be realized, thus being rapid and effective, not only having significant meaningfor early treatment of adolescent idiopathic scoliosis and saving medical cost, but also providing therapeutic targets and important basis for clinical application for gene therapy and drug therapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to ENST00000602322.1 and / or MIR210HG as a molecular marker for juvenile idiopathic scoliosis and its application. Background technique [0002] Idiopathic scoliosis (Idiopathic Scoliosis, IS) is a disease with a high incidence rate (1.3%) and seriously affects the quality of life, accounting for about 80% of the total incidence of scoliosis. According to the age of onset, it is divided into infantile idiopathic scoliosis (Infantile Idiopathic Scoliosis, IIS) (0.3 years old), juvenile idiopathic scoliosis (Juvenile Idiopathic Scoliosis, JIS) (3-9 years old), adolescent idiopathic scoliosis Adolescent Idiopathic Scoliosis (AIS) (10-16 years old). Among them, AIS accounts for about 80% of the patients with idiopathic scoliosis. The etiology of AIS is not yet clear, and there are many hypotheses describing its pathogenesis, but none of them can explain the occurrence of AIS well. [0003...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 王以朋李子全李政垚刘书中王牧川牛潼
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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