Detection method of invasive periodontitis
A technique for periodontitis and detection of people, which is applied in the field of detection of invasive periodontitis, and can solve the problem of not having polymorphism
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example 1
[0054] Example 1 to obtain samples
[0055] Collect a case of patients, children, female, 15 years old, students, excellent academic performance, left-behind children. Dental examination revealed a total of 25 teeth in the mouth. Poor oral hygiene, supragingival calculus (+++), BOP (+), red and swollen gums, probing depth of periodontal pockets 3-7mm, periodontal pus on teeth 35 and 36; bleeding index 3; teeth 15, 22, 26 teeth, 32-36 teeth loose III°, 37 teeth, 46 teeth loose I°, 21 teeth, 31 teeth, 41 teeth missing (see figure 1 ). The deciduous teeth erupted normally without premature loss. CBCT showed that the full-mouth alveolar bone resorption reached 1 / 3 of the root length to the root tip, and the immature tooth germs of teeth 18, 28, 38 and 48 could be seen (see figure 2 ). The patient's physical development was moderate, with no abnormal development of skin, hair, and nails, and normal sweat secretion. The jaw bone development was normal and the facial shape was...
Embodiment 2
[0057] Example 2 sample preparation (using Tiangen BioMedium Blood Genomic DNA Extraction Kit (0.5-3ml) (DP332))
[0058] 1. Sampling: Use EDTA-K2 anticoagulant tubes to draw 5ml of blood from adolescent patients with aggressive periodontitis and their family members.
[0059] 2. Add 200 μl Proteinase K (20 mg / ml) solution to a 15 ml centrifuge tube.
[0060] 3. Processing materials: add 3ml of blood sample, and mix well in the above-mentioned 15ml centrifuge tube filled with Proteinase K solution.
[0061] 3. Add 2.4ml buffer GE to the centrifuge tube containing the blood sample, shake for 30sec and mix well.
[0062] 4. Place at 65°C for 10 minutes, shake once every 3 minutes to facilitate lysis.
[0063] 5. Add 2ml of absolute ethanol to the sample and mix well. At this time, flocculent precipitation may appear.
[0064] 6. Transfer half of the solution and flocculent precipitate obtained in the previous step to an adsorption column CB5 (the adsorption column is placed i...
Embodiment 3
[0070] Example 3 Using the next-generation sequencing technology of human peripheral blood whole exome for clinical verification
[0071] A case of aggressive periodontitis was sampled using the next-generation sequencing technology of the whole exome of human peripheral blood. At the same time, the samples of the father, mother and fraternal twin brother of the aggressive periodontitis case were taken as reference comparisons, using SEQ ID NO: 1-SEQID NO:6 primers amplified three genes ANKRD36C, SREK1IP1 and TMEM97 to analyze their polypeptide properties, and the results showed that there were frameshift mutations in these three genes image 3 , the main transcript numbers and nucleotide / amino acid changes are as follows:
[0072] ENST00000456556:p.Glu491fs / c.1470dupT,
[0073] ENST00000513458:p.Arg91fs / c.271dupA,
[0074] ENST00000226230:p.Ter177fs / c.528dupA.
[0075] ANKRD36C, a T is inserted at position 1470 of the gene CDS, resulting in changes in the amino acids after...
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