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Inter- and intra-test agreement of three commercially available molecular diagnostic tests for the identification of periodontal pathogens

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Abstract

Objective

The aim of the study was to compare the agreement and reproducibility of three test kits (IAI Pado Test 4⋅5®, micro-IDent®, and Carpegen® Perio Diagnostik) regarding the detection of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Aggregatibacter actinomycetemcomitans.

Materials and methods

Periodontal plaque samples from 20 patients suffering from severe chronic or aggressive periodontitis were split into two identical batches. Samples of batch 1 were evaluated immediately. Samples of batch 2 were analyzed after a storage period of 2 weeks at 4 °C.

Results

The conformity of results obtained by the evaluated test kits as well as the concordant detection rate of periodontopathogens proved to be low. The agreement of the detection rate depends on the chosen test kit. The intra-test reproducibility of the qualitative analysis of identical plaque samples was perfect only for Carpegen® Perio Diagnostik. The quantitative analysis of the results expressed as n-fold difference of the test results revealed an only poor inter-test agreement between the test systems.

Conclusions

Results of test kits for the identification of periodontopathogens are significantly influenced by the detection technology applied. The most advanced technology tended to give the most reliable results.

Clinical relevance

As microbiological test results are supposed to be used for the planning of subsequent treatment strategies, clinicians should be aware of the limitations of results obtained by the different test systems and should use the results only in conjunction with clinical data for subsequent therapeutic decisions.

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Notes

  1. Distributed as meridol® Paro Diagnostik by GABA GmbH until 2009.

References

  1. Kinane DF (2001) Causation and pathogenesis of periodontal disease. Periodontol 2000 25:8–20

    Article  PubMed  Google Scholar 

  2. Paster BJ, Boches SK, Galvin JL, Ericson RE, Lau CN, Levanos VA, Sahasrabudhe A, Dewhirst FE (2001) Bacterial diversity in human subgingival plaque. J Bacteriol 183:3770–3783

    Article  PubMed Central  PubMed  Google Scholar 

  3. Pozhitkov AE, Beikler T, Flemmig T, Noble PA (2011) High-throughput methods for analysis of the human oral microbiome. Periodontol 2000 55:70–86

    Article  PubMed  Google Scholar 

  4. Socransky SS, Haffajee AD (1991) Microbial mechanisms in the pathogenesis of destructive periodontal diseases: a critical assessment. J Periodontal Res 26:195–209

    Article  PubMed  Google Scholar 

  5. Loomer PM (2004) Microbiological diagnostic testing in the treatment of periodontal diseases. Periodontol 2000 34:49–56

    Article  PubMed  Google Scholar 

  6. Zambon JJ (1995) American Academy of Periodontology Consensus report periodontal disease. Microbial factors. Ann Periodontol 67:879–925

    Google Scholar 

  7. Ezzo PJ, Cutler CW (2003) Microorganisms as risk indicators for periodontal disease. Periodontol 2000 32:24–35

    Article  PubMed  Google Scholar 

  8. Tonetti MS, Mombelli A (1999) Early onset periodontitis. Ann Periodontol 4:39–52

    Article  PubMed  Google Scholar 

  9. Müller HP, Heinecke A, Borneff M, Kiencke C, Knopf A, Pohl S (1998) Eradication of Actinobacillus actinomycetemcomitans from the oral cavity in adult periodontitis. J Periodontal Res 33:49–58

    Article  PubMed  Google Scholar 

  10. Beikler T, Karch H, Flemming T (2005) Mikrobiologische Diagnostik in der Parodontitistherapie. Gemeinsame Stellungnahme der Deutschen Gesellschaft für Parodontologie (DGP) und der Deutschen Gesellschaft für Zahn-, Mund- und Kieferheilkunde (DGZMK). Dtsch Zahnarztl Z 60:660–662

    Google Scholar 

  11. Mombelli A, Casagni F, Madianos PN (2002) Can presence or absence of periodontal pathogens distinguish between subjects with chronic and aggressive periodontitis? A systematic review. J Clin Periodontol 29(suppl 3):10–21

    Article  PubMed  Google Scholar 

  12. Newman MG, Takei H, Klokkevold PR, Carranza FA (2006) Carranza’s clinical periodontology. Saunders Elsevier, St. Louis, pp 579–601, Chapter 37

    Google Scholar 

  13. Loesche WJ, Lopatin DE, Stoll J, van Poperin N, Hujoel PP (1992) Comparison of various detection methods for periodontopathic bacteria: can culture be considered the primary reference standard? J Clin Microbiol 30:418–426

    PubMed Central  PubMed  Google Scholar 

  14. Savitt ED, Strzempko MN, Vaccaro KK, Peros WJ, French CK (1988) Comparison of cultural methods and DNA probe analyses for the detection of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius in subgingival plaque samples. J Periodontol 59:431–438

    Article  PubMed  Google Scholar 

  15. Kornman KS, Jones A, Nolan P, Alvarado R, Manwell M, Wood R (1992) Culture, DNA probes and mononucleal antibodies for the detection of bacteria in periodontitis. J Dent Res 71:121

    Google Scholar 

  16. Riggio MP, Macfarlane TW, Mackenzie D, Lennon A, Smith AJ, Kinane D (1996) Comparison of polymerase chain reaction and culture methods for detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in subgingival plaque samples. J Periodontal Res 31:496–501

    Article  PubMed  Google Scholar 

  17. Eick S, Pfister W (2002) Comparison of microbial cultivation and a commercial PCR based method for detection of periodontopathogenic species in subgingival plaque samples. J Clin Periodontol 29:638–644

    Article  PubMed  Google Scholar 

  18. Bürklin T, Hassenrück S, Böddinghaus B, Schacher B, Eickholz P, Kim T, Schaecken MT, Renggli HH, Ratka-Krüger P (2002) Vergleich von zwei in der Parodontologie verwendeten Gensondentests - IAI Pado Test 4⋅5® und micro-IDent®. Parodontol 13:235–242

    Google Scholar 

  19. Jervøe-Storm P, Koltzscher M, Falk W, Dörfler A, Jepsen S (2005) Comparison of culture and real-time PCR for detection and quantification of five putative periodontopathogenic bacteria in subgingival plaque samples. J Clin Periodontol 32:778–783

    Article  PubMed  Google Scholar 

  20. Yaskell T, Torresyap G, Teles RP, Socransky SS, Haffajee AD (2007) Comparison of molecular techniques for microbial assessment of subgingival biofilms. J Dent Res 86:2923

    Google Scholar 

  21. Frank P, Eick S, Eickholz P, Kim C, Kim T (2008) A.a.-Nachweis mittels Realtime-PCR und Gensondentest bei Patienten verschiedener ethnischer Herkunft. Parodontol 19:317–359

    Google Scholar 

  22. Cosgarea R, Zehaczek S, Bäumer A, Kim T (2008) Vergleich zweier unterschiedlicher Gensondentests zum Nachweis parodontaler Pathogene. Parodontol 19:317–359

    Google Scholar 

  23. Lang N, Bartold PM, Cullian M, Jeffcoat M, Mombelli A, Murakami S, Page R, Papapanou P, Tonettti M, Van Dyke T (1999) Consensus report: aggressive periodontitis. Ann Periodontol 4:53

    Article  Google Scholar 

  24. Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33:159–174

    Article  PubMed  Google Scholar 

  25. Farztdinov V, McDyer F (2012) Distributional fold change test—a statistical approach for detecting differential expression in microarray experiments. Algorithms Mol Biol 7:29. doi:10.1186/1748-7188-7-29

    Article  PubMed Central  PubMed  Google Scholar 

  26. Moore WEC, Moore LVH (1994) The bacteria of periodontal diseases. Periodontol 2000 5:66–77

    Article  PubMed  Google Scholar 

  27. Krigar DM, Kaltschmitt J, Krieger JK, Eickholz P (2007) Two subgingival plaque sampling strategies used with RNA–probes. J Periodontol 78:72–78

    Article  PubMed  Google Scholar 

  28. Jervøe-Storm P, Alahdab H, Koltzscher M, Fimmers R, Jepsen S (2007) Comparison of curet and paper point sampling of subgingival bacteria as analyzed by real-time polymerase chain reaction. J Periodontol 78:909–917

    Article  PubMed  Google Scholar 

  29. Dannewitz B, Pohl S, Eickholz P, Kim TS (2007) Clinical and microbiological effects of a combined mechanic-antibiotic therapy in subjects with Actinobacillus actinomycetemcomitans-associated periodontitis. Am J Dent 20(3):153–156

    PubMed  Google Scholar 

  30. Slots J, Reynolds HS, Genco RJ (1980) Actinobacillus actinomycetemcomitans in human periodontal disease: a cross sectional microbiological investigation. Infect Immun 29:1013–1020

    PubMed Central  PubMed  Google Scholar 

  31. Zambon JJ, Christersson LA, Slots J (1983) Actinobacillus actinomycetemcomitans in human periodontal disease. Prevalence in patient groups and distribution of biotypes and serotypes within families. J Periodontol 54:707–711

    Article  PubMed  Google Scholar 

  32. Silveira VR, Nogueira MV, Nogueira NA, Lima V, Furlaneto FA, Rego RO (2013) Leukotoxicity of Aggregatibacter actinomycetemcomitans in generalized aggressive periodontitis in Brazilians and their family members. J Appl Oral Sci 21(5):430–436. doi:10.1590/1679-775720130252

    Article  PubMed Central  PubMed  Google Scholar 

  33. Heid CA, Stevens J, Livak KJ, Williams PM (1996) Real time quantitative PCR. Genome Res 6:986–994

    Article  PubMed  Google Scholar 

  34. Lau L, Sanz M, Herrera D, Morillo JM, Martín C, Silva A (2004) Quantitative real-time polymerase chain reaction versus culture: a comparison between two methods for the detection and quantification of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis in subgingival plaque samples. J Clin Periodontol 31(12):1061–1069

    Article  PubMed  Google Scholar 

  35. Boutaga K, van Winkelhoff AJ, Vandenbroucke-Grauls CM, Savelkoul PH (2005) Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR. FEMS Immunol Med Microbiol 45(2):191–199

    Article  PubMed  Google Scholar 

  36. Verner C, Lemaitre P, Daniel A, Giumelli B, Lakhssassi N, Sixou M (2006) Carpegen® real-time polymerase chain reaction vs. anaerobic culture for periodontal pathogen identification. Oral Microbiol Immunol 21:341–346

    Article  PubMed  Google Scholar 

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Acknowledgments

We thank Dr. rer. nat. Birte Holtfreter, Department of Restorative Dentistry, Periodontology, and Endodontology, Ernst-Moritz-Arndt University Greifswald, for the statistical analysis of the study data.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Private Dental Practice, Bahnhofstr. 9, 63654, Buedingen, Germany

    Martin Untch

  2. Department of Periodontology, University Hospital Wuerzburg, Pleicherwall 2, 97070, Wuerzburg, Germany

    Ulrich Schlagenhauf

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  1. Martin Untch
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  2. Ulrich Schlagenhauf
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Correspondence to Ulrich Schlagenhauf.

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Untch, M., Schlagenhauf, U. Inter- and intra-test agreement of three commercially available molecular diagnostic tests for the identification of periodontal pathogens. Clin Oral Invest 19, 2045–2052 (2015). https://doi.org/10.1007/s00784-015-1418-3

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  • DOI: https://doi.org/10.1007/s00784-015-1418-3

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