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Early screening method and kit of ankylosing spondylitis susceptibility gene
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A technology for ankylosing spondylitis and susceptibility genes, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc.
Active Publication Date: 2021-06-25
北京科力丹迪生物医疗科技有限公司
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At present, there is no very effective early detection method in clinic, so it is urgent to establish a highly sensitive, economical and simple molecular technology screening method to promote the early screening of ankylosing spondylitis
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Embodiment 1
[0101] Feasibility analysis of SNP site screening of human ankylosing spondylitis susceptibility-related genes
[0102] The inventors screened and evaluated the occurrence-related sites of ankylosing spondylitis that were verified in large-scale pathological control group clinical studies by searching NCBI's domestic and foreign genome-wide association study (GWAS). Selected 14 single nucleotide polymorphism sites that are significantly related to the risk of ankylosing spondylitis in the Chinese population, and are independent of each other, there is no linkage disequilibrium, so the site selection of the present invention is representative, independent and Risk values are cumulative and can be used to assess an individual's risk of developing ankylosing spondylitis.
[0106] System validation includes accuracy, specificity, sensitivity, precision, and comparison among personnel.
[0107] Accuracy verification scheme: 20 cases were detected at each site, compared with Sanger sequencing, the expected target was 95%.
[0108] Specificity Validation Protocol: Included in Accuracy, expected target 95%.
[0109] Sensitive verification scheme: using human genomic DNA positive samples as templates, the DNA contents of calibration samples were 1ng / μL, 5ng / μL, 10ng / μL, 50ng / μL, and 100ng / μL for sensitivity inspection.
[0110] The precision verification plan (including intra-batch, inter-batch, and personnel comparisons, not involving inter-instrument comparisons) has an expected target of 95%.
[0111] Intra-assay precision: The same batch of each sample was repeated 3 times to compare the intra-assay precision.
[0112] Inter-batch precision: The same operator tests the same sample in multiple batches to...
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Abstract
The invention provides an early screening method of ankylosing spondylitis susceptibility genes and a kit, and particularly, the invention takes differences between gene spectrums of Chinese people and European and American people into consideration, and screens out a combination of SNP loci of ankylosing spondylitis susceptibility related genes of Chinese people; a nucleic acidmassspectrometer is used for wide (high-throughput detection sites and high-throughput detection samples) screening and inspection of genetic markers related to ankylosing spondylitis. The method disclosed by the invention is high in detection success rate, good in technical reproducibility and high in cost performance, can realize multi-gene detection of a single small sample, and meets maximum use of the small sample; the method has the technical advantages of high accuracy and high sensitivity, the detection result is stable, and the detection positive rate is increased.
Description
technical field [0001] The invention belongs to the field of biological technology, in particular, the invention relates to an early screening method and kit for susceptibility genes of ankylosing spondylitis. Background technique [0002] Ankylosing spondylitis (AS) is a disease characterized by inflammation of the sacroiliac joints and spinal attachment points. Five years after the onset of ankylosing spondylitis, about 4% of the patients lost part of their working ability, and about 50% of the patients lost their working ability after 45 years of onset. Genetic factors play an important role in the pathogenesis of AS. It is reported abroad that the incidence of ankylosing spondylitis is 0.1% in the general population, while the prevalence in relatives of patients with ankylosing spondylitis is as high as 4.0%, which is 40 times higher than the general population. AS is a polygenic disease with a complex genetic background, so we need to conduct multi-level three-dimensi...
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