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Probe and primer for detecting single nucleotide polymorphism related to chronic periodontitis, and kit thereof

A single nucleotide polymorphism, chronic periodontitis technology, applied in the biological field, can solve the problem that the susceptibility of chronic periodontitis is blank

Active Publication Date: 2013-06-05
上海市口腔病防治院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is still blank to use single nucleotide polymorphisms to assist clinical diagnosis of chronic periodontitis susceptibility

Method used

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  • Probe and primer for detecting single nucleotide polymorphism related to chronic periodontitis, and kit thereof
  • Probe and primer for detecting single nucleotide polymorphism related to chronic periodontitis, and kit thereof
  • Probe and primer for detecting single nucleotide polymorphism related to chronic periodontitis, and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] The assembly of embodiment 1 kit

[0131] Kit preparation:

[0132] Step 1, substrate processing and sampling

[0133] Wash the slides with double distilled water, soak them overnight in lye, take them out, wash them three times with double distilled water, then soak them in HCl with a volume fraction of 1% for 30 minutes, and put them in a slide tank after cleaning. use. The cleaned glass slides were aminated with 2% 4-aminobutyltriethoxysilane aqueous solution by volume, and then subjected to isothiocyanation treatment with 1mmol / L benzene isothiocyanate aqueous solution, and blown with nitrogen. After drying, store at 4°C in the dark.

[0134] Chip Zip probes are randomly combined oligonucleotide fragments with a length of 24bp. These oligonucleotide sequences are compared with the target genome sequence using the BLAST function on the NCBI website. The least derived set (as Zip probe sequences).

[0135] Zip probe for rs2891168 wild type: 5'-GCAAGTCTACCTAATCTCTGA...

Embodiment 2

[0146] The detection of embodiment 2 kit

[0147] Step 1, sample preparation

[0148] Saliva samples of each case were prepared, and genomic DNA was extracted from oral exfoliated cells of saliva by conventional phenol-chloroform method.

[0149] Step 2, PCR amplification

[0150] Using the genomic DNA of each sample as a template, PCR amplification was carried out with the specific amplification primer pair of rs2891168 and the specific amplification primer pair of rs1800796 respectively.

[0151] PCR reaction components (15μl system): 20ng genomic DNA, 2×Taq PCR Master Mix, 0.4μM each primer, RNase-free water complement.

[0152] The reaction conditions of Touch-down PCR are: start at 94°C for 5min, denature at 94°C for 30s, anneal at 64°C for 30s (down 0.5°C per cycle), extend at 72°C for 1min, cycle 14 times; denature at 94°C for 30s, anneal at 57°C for 30s , extended at 72°C for 1 min, cycled 30 times, and extended at 72°C for 10 min.

[0153] After the reaction was c...

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Abstract

The invention provides a probe and a primer for detecting single nucleotide polymorphism related to chronic periodontitis, and a kit thereof. The probe and the primer for detecting single nucleotide polymorphism related to chronic periodontitis disclosed by the invention comprise specific amplification primer pairs at single nucleotide polymorphic site rs2891168, specific amplification primer pairs at single nucleotide polymorphic site rs1800796, a detection probe of the single nucleotide polymorphic site rs2891168, and a detection probe of the single nucleotide polymorphic site rs1800796. A kit prepared by the probe and the primer for detecting the single nucleotide polymorphism related to the chronic periodontitis is also further provided by the invention. The probe, the primer and the kit disclosed by the invention are especially suitable for single nucleotide polymorphism research of chronic periodontitis of Chinese Han nationality population.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe, a primer, a chip and a kit for detecting chronic periodontitis-related single nucleotide polymorphisms. Background technique [0002] Chronic periodontitis is the most common disease among periodontal diseases, and it is also one of the refractory periodontal diseases. The traditional diagnostic method of chronic periodontitis is mainly based on the clinical signs, and the diagnosis, classification and evaluation of chronic periodontitis are carried out by measuring the periodontal index, the degree of alveolar bone attachment loss, the degree of tooth looseness, and the depth of periodontal pockets. Type, and according to the severity of each index, determine the development stage of periodontitis, and implement different types of treatment. This diagnostic method is intuitive, simple, and easy to master. However, it is difficult to diagnose early periodontitis because it ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 陈栋汪黎明张洁冯坤
Owner 上海市口腔病防治院
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