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A method for detection of susceptibility gene of congenital edentulousness

A susceptibility gene and congenital technology, applied in the field of gene chips, can solve the problems of large amount of sequencing results, high price, time-consuming, etc., and achieve the effect of enriching the spectrum of pathogenic mutations, expanding the coverage of diagnosis, and facilitating the interaction

Active Publication Date: 2019-09-10
PEKING UNIV SCHOOL OF STOMATOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Sanger sequencing is time-consuming, consumable, and manpower-intensive; exome sequencing and whole-genome sequencing are not only expensive, but also result in huge amounts of data, long cycles, and high costs

Method used

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  • A method for detection of susceptibility gene of congenital edentulousness
  • A method for detection of susceptibility gene of congenital edentulousness
  • A method for detection of susceptibility gene of congenital edentulousness

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Acquisition of DNA samples to be sequenced

[0028] 1) Collect 2 mL of saliva samples from patients, family members and normal controls, and collect them in the saliva sample collection tubes in the Oragene DNA (OG-500) DNA collection kit (DNA Genoteck, general agent of Bipi Biotechnology Co., Ltd.), at room temperature Save it.

[0029] 2) Use the Oragene·DNA (OG-500) DNA collection kit (DNA Genoteck, the general agent of Bipi Biotechnology Co., Ltd.) to extract human genomic DNA from the collected saliva samples.

[0030] 3) 1 μl of the extracted DNA was loaded as a sample, and the amount of genomic DNA was checked by 1% agarose gel electrophoresis, and the DNA was quantified with a UV-640 Beckman spectrophotometer. Aliquot with appropriate volume and freeze at -20°C.

Embodiment 2

[0031] The screening of embodiment 2 congenital edentulous gene

[0032] Collect the reported genes that may be related to congenital edentulousness, according to the importance of gene expression in the development of tooth germ, the frequency of occurrence in various literatures or reports, data and other factors, from more than 60 congenital edentulous 35 edentulous candidate genes were screened out, and classified according to whether mutations were detected in human congenital edentulous patients, as shown in Table 1:

[0033] Table 1: Selected congenital edentulous genes

[0034]

[0035]

[0036] The test samples used in the detection method of congenital edentulous susceptibility gene of the present invention come from patients, family members and normal controls, and can detect the gene mutations in the 35 gene sequences in Table 1 of congenital edentulous patients On the one hand, use clinical big data to verify the existing research and development results, a...

Embodiment 3

[0038] 1. Reagents for detection

[0039] Library Construction Reagents:

[0040] (1) Primer library (synthesized by Life Technologie)

[0041] (2) Ion AmpliSeq TM Library Kits 2.0(Life Technologies)

[0042] (3)Ion Xpress TM Barcode Adapters (Life Technologies)

[0043] (4) XP Reagent (Beckman Coulter)

[0044](5) Agilent High Sensitivity DNA Kit (Agilent)

[0045] Ion OneTouch TM Reagent: Ion PGM TM Hi-Q TM OT2Kit (Life Technologies)

[0046] Ion Torrent sequencing reagent: Ion PGM TM Hi-Q TM Sequencing Kit (Life Technologies)

[0047] 2. Experimental process

[0048] (1a) Use two sets of primer libraries (see Example 7 for the sequences of the primer libraries) to amplify the full-length gene in the target region to obtain preliminary sequencing library fragments. The amplification system and PCR amplification conditions are as follows:

[0049] Table 2: Amplification System

[0050] Element Amount added AmpliSeq TM HiFi Mix(red cap)

...

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Abstract

Disclosed are congenital absence susceptibility gene detection chip and detection method. By the detection chip and the detection method, congenital absence genetic diagnosis efficiency can be improved greatly, diagnosis coverage of candidate genes can be expanded, a pathogenic mutant spectrum of congenital absence can be enriched greatly, analysis on interactive action between genes is facilitated, better understanding on the pathogenic factor of the disease can be obtained, and the high-value research results brings good to genetic counseling and antenatal diagnosis of congenital absence.

Description

technical field [0001] The invention relates to a gene chip, in particular to a method for detecting susceptibility genes of congenital tooth loss. Background technique [0002] A congenital reduction in the number of teeth due to abnormal development of tooth germs is called congenital edentulous. In recent years, with the localization of various molecules in tooth development, such as transfer factors, growth factors, receptor molecules, cell surface molecules, intercellular molecules, etc., and the study of their regulation during tooth development, the innate The research on the genetic factors of sexual edentulousness is more in-depth, but the views of various scholars are inconsistent. For example, Vastardis et al. found that when the MXS1 gene locus is mutated, mice can show severe tooth loss; humans can show congenital loss of second premolars and third molars. In addition, some scholars have found that epidermal growth factor (Epideral growth factor, EGF), epiderm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6869C12Q1/6837
Inventor 韩冬冯海兰刘洋刘浩辰王升威王越刘湘宋晓玲
Owner PEKING UNIV SCHOOL OF STOMATOLOGY
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