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Single nucleotide polymorphism of human NOS3 gene 64 th locus and application of single nucleotide polymorphism

A single nucleotide polymorphism, 64-bit technology, applied in the fields of molecular biology and medicine, can solve the problems of immune dysfunction, systematic research on RA single nucleotide polymorphism, breaking autoimmune tolerance, etc.

Inactive Publication Date: 2017-12-15
王思捷
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the pathogenesis of RA has not yet been fully elucidated, it is generally believed that the occurrence of RA is caused by the stimulation of various risk factors by individuals with genetic susceptibility to induce immune dysfunction and break autoimmune tolerance.
At present, there is no systematic study of RA single nucleotide polymorphism in my country

Method used

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  • Single nucleotide polymorphism of human NOS3 gene 64 th locus and application of single nucleotide polymorphism
  • Single nucleotide polymorphism of human NOS3 gene 64 th locus and application of single nucleotide polymorphism
  • Single nucleotide polymorphism of human NOS3 gene 64 th locus and application of single nucleotide polymorphism

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 is the sequence determination of exon 1 of NOS3 gene in 8 RA family populations.

[0017] (1) RA family enrollment and sample collection

[0018] In this study, clinically diagnosed RA patients were selected, and families with more than two RA patients were screened by inquiring about their family history of RA, and all RA patients and at least two non-RA healthy members in the family were selected. In the case of obtaining the informed consent of relevant family members of such RA family members, check representative family members other than patients to confirm that they are indeed RA patients and healthy people who are not RA, where healthy people especially refer to non-RA patients. The research subjects were all from Fujian Province and its surrounding areas, and a total of 8 eligible RA families were collected. The clinical and pathological data of RA patients were collected, and 5ml of fasting venous anticoagulant blood was collected from all of them. ...

Embodiment 2

[0028] Example 2 is to use the real-time fluorescent quantitative PCR detection method of NOS3 gene 64G / A nucleotide polymorphism constructed in the present invention to verify in population samples.

[0029] In addition, 100 clinically diagnosed RA patients different from Example 1 and 250 healthy people were randomly selected, and 2 ml of fasting venous anticoagulant blood samples were taken from each person with informed consent, and the real-time fluorescent quantitative PCR detection reagent constructed by the present invention was used. The box and method are used to detect the 64G / A nucleotide polymorphism of the NOS3 gene. The specific operations and parameters are the same as in Example 1. For the amplification curve of healthy people, see Figure 4 , RA patients see image 3 and Figure 4 . The results are shown in Table 2. The 64th position in healthy people is all G nucleotides (100%); while the 64th position in RA patients is only 82% ( Figure 4 ), as high as...

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Abstract

Revelation of genetic susceptibility gene loci is an important direction of studying rheumatoid arthritis currently. The invention provides G / A nucleotide polymorphism of a human NOS3 gene 64 th locus, the susceptibility sequence of the locus is shown in SEQ ID NO.1, through the study of two aspects of families of rheumatoid arthritis and crowds of rheumatoid arthritis, the relation of the locus and the rheumatoid arthritis susceptibility is revealed, on this basis, a kit for conducting real-time fluorescence quantification detection on the sensitive specificity of the locus is constructed, the kit includes specific primers SEQ ID NO.3 and SEQ ID NO.4, and TaqMan probes SEQ ID NO.5 and SEQ ID NO.6, a real-time fluorescence quantification PCR system and a condition construction and detection method are optimized, and the methods can provide a basis for susceptibility mechanism analysis, susceptibility gene detection and accurate diagnosis of Chinese crowds suffering from rheumatoid arthritis.

Description

technical field [0001] The invention belongs to the fields of molecular biology and medicine, and in particular relates to the detection of the single nucleotide polymorphism at the 64th position of the human NOS3 gene and its correlation with rheumatoid arthritis; the invention also relates to the real-time fluorescence quantification of the site PCR detection kits and methods. Background technique [0002] Rheumatoid arthritis (Rheumatoid Arthritis, RA) is a systemic autoimmune disease characterized by joint inflammation, the main pathological changes are chronic synovitis and vasculitis, when the articular cartilage and bone are involved, Joint deformity and ankylosis gradually develop, leading to loss of joint function. Although the pathogenesis of RA has not been fully elucidated so far, it is generally believed that the occurrence of RA is caused by individuals with genetic susceptibility stimulated by various risk factors to induce immune dysfunction and break autoim...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/156C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 王思捷
Owner 王思捷