Search

KR-102962019-B1 - Composition for detecting oral bacteria helpful in diagnosing periodontitis and uses thereof

KR102962019B1KR 102962019 B1KR102962019 B1KR 102962019B1KR-102962019-B1

Abstract

The present invention relates to a composition for detecting oral bacteria that can aid in the diagnosis of periodontitis, comprising a primer set capable of detecting bacteria causing periodontitis, a kit using the same, or a method for providing information. The primer set of the present invention can diagnose periodontitis by detecting four types of oral bacteria. In particular, the primer set of the present invention can specifically diagnose only periodontitis among various periodontal diseases including gingivitis, and can be effectively utilized in compositions, kits, or information provision methods for detecting oral bacteria.

Inventors

  • 허대욱
  • 안인숙
  • 김형록
  • 이연희
  • 홍지연
  • 이병진
  • 노승희

Assignees

  • 주식회사 덴오믹스

Dates

Publication Date
20260508
Application Date
20230728

Claims (10)

  1. A first primer set comprising a primer having the nucleotide sequence of SEQ ID NO. 1, a primer pair having the nucleotide sequence of SEQ ID NO. 2, and a probe having the nucleotide sequence of SEQ ID NO. 3; A second primer set comprising a primer having the nucleotide sequence of SEQ ID NO. 4, a primer pair having the nucleotide sequence of SEQ ID NO. 5, and a probe having the nucleotide sequence of SEQ ID NO. 6; A third primer set comprising a primer having the nucleotide sequence of SEQ ID NO. 7, a primer pair having the nucleotide sequence of SEQ ID NO. 8, and a probe having the nucleotide sequence of SEQ ID NO. 9; and A fourth primer set comprising a primer having the nucleotide sequence of SEQ ID NO. 10, a primer pair having the nucleotide sequence of SEQ ID NO. 11, and a probe having the nucleotide sequence of SEQ ID NO. 12; A composition for detecting oral bacteria causing periodontitis that includes all of the A composition characterized in that the oral bacteria are Porphyromonas gingivalis , Campylobacter rectus, Prevotella nigrescens , and Streptococcus mutans .
  2. delete
  3. delete
  4. delete
  5. A composition according to claim 1, characterized in that the primer set is used in PCR (polymerase chain reaction).
  6. In claim 5, the above PCR is quantitative PCR (qPCR), real-time PCR, reverse transcription PCR (RT-PCR), solid phase PCR, competitive PCR, overlap-extension PCR, multiplex PCR, nested PCR, inverse PCR, ligation-mediated PCR, intersequence-specific PCR (ISSR), methylation-specific PCR (MSP), colony PCR, miniprimer PCR, nanoPCR (nanoparticle-assisted PCR), thermal asymmetric interlaced PCR (TAIL-PCR), touchdown (step-down) PCR, hot start PCR, in silico PCR, allele-specific A composition characterized by being one or more selected from the group consisting of allele-specific PCR, assembly PCR, asymmetric PCR, dial-out PCR, digital PCR (dPCR), and helicase-dependent amplification.
  7. A kit for detecting oral bacteria comprising the composition of claim 1.
  8. i) a step of extracting DNA from a biological sample isolated from an individual suspected of having periodontitis; and ii) a step of detecting bacteria using the DNA obtained in step i) and the composition of claim 1; A method for providing information for the diagnosis of periodontitis including, A method for providing information characterized in that the bacteria in step ii) above are Porphyromonas gingivalis , Campylobacter rectus, Prevotella nigrescens , and Streptococcus mutans .
  9. A method for providing information according to claim 8, wherein the biological sample in step i) is an oral rinse and/or saliva.
  10. delete

Description

Composition for detecting oral bacteria helpful in diagnosing periodontitis and uses thereof The present invention relates to a composition for detecting oral bacteria that can aid in the diagnosis of periodontitis, comprising a primer set capable of detecting bacteria causing periodontitis, a kit using the same, or a method for providing information. Periodontal disease, commonly referred to as periodontitis, is a complex, chronic oral inflammatory disease that destroys the soft and hard tissues supporting the teeth. If left untreated, the alveolar bone surrounding the teeth is lost, causing the teeth to loosen and subsequently leading to tooth loss. Major symptoms include inflammation, gum bleeding, gum recession, periodontal pockets between the teeth and gums, and, in cases of severe periodontitis, looseness or damage to the teeth. One of the current diagnostic methods for periodontitis is the method of measuring periodontal pocket depth by inserting a probe into the sulcus; this is the most fundamental diagnostic method as it confirms the extent of alveolar bone loss and assesses the degree of gingival inflammation. However, the method of measuring periodontal pocket depth can introduce errors depending on the shape of the teeth and the degree of gingival inflammation. Confirming bone loss on radiographs is, along with the periodontal pocket probe, one of the most basic diagnostic methods for periodontitis. While this method can show alveolar bone loss on the mesiodistal surfaces of the teeth using two-dimensional imaging, it has limitations in that it cannot show alveolar bone loss on the buccal and lingual surfaces of the teeth, which overlap with the image. Furthermore, the aforementioned method of confirming periodontal pocket depth and bone loss on radiographs has a problem in that it only shows the results of alveolar bone loss (loss of attachment) due to the progression of periodontitis prior to the time of diagnosis, and thus cannot show the current active state of the disease. In addition, the most useful method currently available to determine whether there is inflammation in the gingiva at the time of diagnosis is to insert a probe into the sulcus between the tooth and the gingiva to check for bleeding. However, this method has limitations in that it is highly likely to produce a false positive result, as there may not actually be inflammation in the gingiva even if bleeding is detected during probing. These conventional methods, which are measured by specialists in the dental office, are cumbersome, time-consuming, and costly, and depending on the method, inevitably entail patient pain. Furthermore, while the measurement of periodontal pockets and clinical attachment levels, along with the observation of alveolar bone levels on radiographs, demonstrate the clinical consequences of periodontal disease, they do not provide information on whether the gum condition of individual teeth is currently causing attachment loss. Periodontal disease, a major cause of tooth loss, is primarily caused by bacterial infection. Therefore, it is crucial to reduce the number of bacteria in the oral cavity and provide active treatment through early diagnosis. Accordingly, there is a growing need to develop a method that can easily diagnose whether the current gum condition of individual teeth is healthy and free of inflammation or if it is in a state of periodontitis, particularly for periodontal disease cases where symptoms are delayed. Figure 1 shows oral and radiographic images of patients with normal periodontitis, gingivitis, and periodontitis. Figure 2a shows the standard curve for quantifying 4 of the 16 types of oral bacteria ( Aggregatibacter actinomycetemcomitans , Filifactor alocis , Porphyromonas gingivalis , Streptococcus mutans ). Figure 2b shows a standard curve for the quantification of 4 types ( Campylobacter rectus , Fusobacterium nucleatum , Porphyromonas endodontalis , Streptococcus sobrinus ) out of 16 types of oral bacteria. Figure 2c shows a standard curve for the quantification of 4 types ( Eikenella corrodens , Lacticaseibacillus casei , Prevotella intermedia , Treponema denticola ) out of 16 types of oral bacteria. Figure 2d shows a standard curve for the quantification of 4 types ( Eubacterium nodatum , Parvimonas micra , Prevotella nigrescens , Tannerella forsythia ) out of 16 types of oral bacteria. Figure 3a shows the real-time PCR results for Aggregatibacter actinomycetemcomitans in saliva: A: Results of the entire patient group, B: Results of the normal periodontal group, C: Results of the gingivitis group, D: Results of the periodontitis group. Figure 3b shows the real-time PCR results for Campylobacter rectus in saliva: A: Results of the entire patient group, B: Results of the normal periodontal group, C: Results of the gingivitis group, D: Results of the periodontitis group. Figure 3c shows the real-time PCR results for Eikenella corrodens in saliva: A: Results of the entire pa