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Cariogenic bacteria quantitative detection method

A quantitative detection method and cariogenic bacteria technology, which is applied in the field of clinical stomatology, can solve the problem of no obvious correlation between carious actinomycetes and achieve the effect of avoiding PCR pollution

Inactive Publication Date: 2016-04-13
STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, real-time fluorescence quantitative PCR was used to detect the content of Actinomycetes naesli and actinomyces caries in saliva samples of different caries-sensitive children, and it was found that the level of Actinomyces naesli in saliva was highly correlated with caries in children. Actinomycetes caries had no significant correlation

Method used

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  • Cariogenic bacteria quantitative detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0022] The count measurement of caries bacteria consistent with the embodiment

[0023] The target population (150 people in total, including 72 males and 78 females, with an average age of 36.2 years) was used in this dental caries risk assessment model. Two dental professional physicians served as inspectors for oral examination and plaque collection, and relevant data were collected on site.

[0024] Wet sterile dry filter paper (1cm×1cm) or cotton swab in sterile wetting solution, the wetting solution is composed of the following components in mass percentage: peptone 0.1%, surfactant 0.1%, sodium chloride 0.85% , the balance is water;

[0025] Use sterile tweezers to take out the wetted filter paper and spread it over the plaque on the buccal surface of the test subject, and stop for 30 seconds to take a sample. Or scrape the surface of plaque on the buccal surface of the teeth with a cotton swab.

[0026] Contact surface microorganism detection method, the steps are as...

Embodiment 2

[0034] A PCR reaction system is designed for each bacterial species, and probes corresponding to the corresponding bacterial species are provided in each PCR reaction tube.

[0035]

[0036] Synthesize the template sequence corresponding to the genotype of the detected strain first, connect the primers to the template sequence, and then use the RNAntpmix labeled with digoxigenin to label the synthesized sequence, so as to complete the probe.

[0037] The DNA of four cariogenic bacteria extracted according to the TIANampBacteriaDNAKit kit was put into the corresponding PCR reaction system, and the reaction system of each strain was established in a single PCR reaction tube; PCR amplification was performed, and the corresponding amplification was recorded Curve; compare the obtained amplification curve with the standard curve, and calculate the template quantity of the sample. Simultaneously record the ratio values ​​of the four bacterial species this time, and combine them w...

Embodiment 3

[0039] Embodiment 3 Combined with the ratio of cariogenic bacteria, draw the corresponding dental caries risk assessment table (LogC is based on 10)

[0040] Table 1 Risk Assessment Form

[0041]

[0042]

[0043] Recent evaluation results:

[0044] The caries risk assessment model was used to assess the caries risk of the target population (a total of 150 people, including 72 males and 78 females, with an average age of 36.2 years). There were 101 low- and medium-risk people and 49 high-risk people. Longitudinal The study conducted a follow-up review after 6 months and 12 months respectively to evaluate the actual new caries of the target population at 6 months and 12 months, and evaluate the effectiveness of this caries risk assessment form. The result is as follows:

[0045]

[0046]

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Abstract

The invention relates to clinical stomatology, in particular to a cariogenic bacteria quantitative detection method which includes the steps of A and B, sampling and preparing a treatment fluid; C, counting strains; D, extracting 1mL of treatment fluid, centrifugally treating the treatment fluid by 12000 r / m for one minute, discarding supernatant, extracting DNA of four types of cariogenic bacteria in the sampled fluid by means of a DNA kit; E, feeding the DNA of four types of cariogenic bacteria into corresponding single-tube PCR (Polymerase Chain Reaction) systems to perform PCR amplification and then recording corresponding amplification curves, comparing the obtained amplification curves with a standard curve to calculate template quantity of the sample; and F, calculating proportion values of the four types of strains and then calculating quantity of different strains within 1cm*1cm. Since proportion of the four types of cariogenic bacteria in dental plaque is detected and according to comparison between the proportion to actual situation of dental caries of people under test, current caries risk assessment model is improved further and becomes more scientific and accurate.

Description

technical field [0001] The invention relates to clinical stomatology, in particular to a quantitative detection method for cariogenic bacteria. Background technique [0002] Caries is a chronic infectious disease, and children are one of the main sensitive populations. Because of the special age stage, the distribution of caries is uneven, which can involve multiple deciduous teeth and cause extensive caries, which in turn affects oral hygiene and physical and mental health. Studies have shown that among Streptococcimutants, Streptococcus mutans and Streptococcus distal are closely related to caries; Lactobacillus plays an important role in the development of caries in young children. In recent years, it has been found that there is a certain correlation between Actinomyces naesi and caries in children. In the prior art, real-time fluorescence quantitative PCR was used to detect the content of Actinomycetes naesli and actinomyces caries in saliva samples of different carie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12Q1/06C12R1/46C12R1/23
CPCC12Q1/06C12Q1/14C12Q1/6851C12Q2531/113
Inventor 杨德琴罗凌飞窦磊刘鑫徐智云黄兴薛慧高敬
Owner STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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