实时荧光定量PCR检测牙周炎患者龈下菌斑的分布

Abstract

Abstract: Objective Realtime quantitative PCR technique would be used to investigate the distribution and levels of A. Actinomycetemcomitans (Aa) and P. Gingivalis (Pg) in subgingival plaque samples from subjects with aggressive periodontitis (AgP) and chronic periodontitis (CP). Methods Subgingival plaque samples were taken from 32 patients with AP,33 patients with CP and 32 periodontal healthy controls. We constructed recombinant plasmid containing speciesspecific gene fragments and the serial dilution of the plasmid standards,and then the bacteria were quantified by realtime PCR with TaqManMGB probes. Results This method showed accurate specificity and sensitivity,and the prevalence of Aa in AgP group was significantly higher than that in CP group,but no significant differences in the counts of both pathogens were found between periodontitis groups. The prevalence and bacterial counts of Pg were significantly higher than those of Aa within each periodontitis group(P＜0.001). In addition,a significant positive correlation was found between the probing depth and the counts of Aa in AgP group (P＜0.01) as well as that of Pg in CP group (P＜0.001). Conclusions These findings indicate the possible relation between the prevalence of Aa in the subgingival plaque and the diagnosis of periodontitis. Moreover,Aa may not be the predominant periodontopathic bacteria in individuals with AgP in Chinese. There is a great potential in the application of the realtime quantitative PCR in the periodontal research.

Key words: aggressive periodontitis, chronic periodontitis, A. actinomycetemcomitans, P. gingivalis, realtime quantitative PCR

CLC Number:

R781.4+2

References

[1]Armitage GC.Development of a classification system for periodontal diseases and conditions[J].Ann periodontol,1999,4(1):1-6
[2]Darveau RP,Tanner A,Page RC.The microbial challenge in periodontitis[J].Periodontol 200,1997,14(1):12-32
[3]Research SaTCotAAoP.Epidemiology of periodontal diseases[J].J Periodontol,2005,76(8):1406-1419
[4]Kuboniwa M,Amano A,Kimura KR,et al.Quantitative detection of periodontal pathogens using realtime polymerase chain reaction with Taqman probes[J].Oral Microbiol Immunol,2004,19(3):168-176
[5]Nonnenmacher C,Dalpke A,Rochon J,et al.Real Time polymerase chain reaction for detection and quantification of bacteria in periodontal patients[J].J Period,2005,76(9):1542-1549
[6]JervoeStorm PM,Koltzscher M,Falk W,et al.Comparison of culture and realtime PCR for detection and quantification of five putative periodontopathogenic bacteria in subgingival plaque samples[J].J Clin Period,2005,32(7):778-783
[7]JervoeStorm PM,Alahdab H,Koltzscher M,et al.Comparison of curet and paper point sampling of subgingival bacteria as analyzed by realtime polymerase chain reaction[J].J Period,2007,78(5):909-917
[8]Goncharoff P,Figurski DH,Stevens RH,et al.Identification of Actinobacillus actinomycetemcomitans: polymerase chain reaction amplification of lktAspecific sequences[J].Oral Microbiol Immunol,1993,8(2):105-110
[9]Leung WK,Ngai VK,Yau JY,et al.Characterization of Actinobacillus actinomycetemcomitans isolated from young Chinese aggressive periodontitis patients[J].J Period Res,2005,40(3):258-268
[10]Kaner D,Christan C,Dietrich T,et al.Timing affects the clinical outcome of adjunctive systemic antibiotic therapy for generalized aggressive periodontitis[J].J Period,2007,78(7):1201-1208
[11]Tomita S,KomiyaIto A,Imamura K,et al.Prevalence of Aggregatibacter actinomycetemcomitans,Porphyromonas gingivalis and Tannerella forsythia in Japanese patients with generalized chronic and aggressive periodontitis[J].Microb Pathog,2013,61/62:11-15
[12]Mombelli A,Casagni F,Madianos PN.Can presence or absence of periodontal pathogens distinguish between subjects with chronic and aggressive periodontitis ? A systematic review[J].J Clin Periodontol,2002,29(s3):10-21
[13]Cortelli JR,Cortelli SC,Jordan S,et al.Prevalence of periodontal pathogens in Brazilians with aggressive or chronic periodontitis[J].J Clin Periodontol,2005,32(8):860-866
[14]Heller D,SilvaBoghossian CM,do Souto RM,et al.Subgingival microbial profiles of generalized aggressive and chronic periodontal diseases[J].Arch Oral Biol,2012,57(7):973-980
[15]Takeuchi Y,Umeda M,Ishizuka M,et al.Prevalence of periodontopathic bacteria in aggressive periodontitis patients in a Japanese population[J].J Period,2003,74(10):1460-1469
[16]Kawada M,Yoshida A,Suzuki N,et al.Prevalence of Porphyromonas gingivalis in relation to periodontal status assessed by real‐time PCR[J].Oral Microbiol Immunol,2004,19(5):289-292

Realtime quantitative PCR for distribution of subgingival microbiota in periodontitis patients

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	. Research progress of matrix metalloproteinases as modulators of periodontitis [J]. , 2019, 39(3): 271-274.
[2]	. Analysis of inhibitory effects of three endodontic irrigations on the pathogens in root canals [J]. , 2019, 39(1): 25-29.
[3]	. Research on the present situation of interplay of type 2 diabetes mellitus, chronic periodontitis and non-surgical periodontal treatment [J]. , 2018, 38(9): 839-843.
[4]	. The relationship between Treponema denticola and periodontal pocket sulfide levels in chronic periodontitis [J]. , 2018, 38(9): 791-794.
[5]	. Accuracy of digital impression in gingival soft tissues of patients with chronic periodontitis [J]. , 2018, 38(5): 422-427.
[6]	. Research progress in the treatment of aggressive periodontitis [J]. , 2018, 38(3): 281-284.
[7]	. Effects of diode laser on the treatment for chronic periodontitis patients with type 2 diabetes mellitus [J]. , 2018, 38(3): 237-240.
[8]	. A study on the association between RAGE gene polymorphisms and type2 diabetes mellitus with chronic periodontitis. [J]. , 2018, 38(1): 33-38.
[9]	. Research progress of the correlation between chronic periodontal disease and chronic kidney disease (CKD) [J]. , 2018, 38(1): 87-91.
[10]	. Application of CBCT in evaluation of alveolar bone changes before and after periodontal therapy in chronic periodontitis [J]. , 2017, 37(5): 444-448.
[11]	. Research progress on the relation between chronic periodontitis and Alzheimer disease [J]. , 2017, 37(4): 364-368.
[12]	. Evaluation of the short term prognosis of implant patients with chronic periodontitis using whole unstimulated saliva [J]. , 2017, 37(4): 350-355.
[13]	. A chronic periodontitis patient with type 2 diabetes mellitus who lost follow up for 7 years after periodontal non-surgical treatment: A case analysis [J]. , 2017, 37(2): 153-157.
[14]	. Changes of the expression levels of IL-23 and IL-17 in GCF of patients with chronic periodontitis before and after treatment [J]. , 2016, 36(6): 536-538.
[15]	. Expression of MMP-2-tryptase double positive mast cells in human chronic periodontitis [J]. , 2016, 36(5): 411-415.