The PadoTest® is the microbiological test that recognises key periodontopathogenic germs including the Aa serotypesand recommends targeted measures for more successful treatment. Dentists and dental hygienists throughout Europe have been using the PadoTest® successfully for years.
Every PadoTest® analysisincludes the determination of the PadoSero serotype. PadoSero is the microbiological test that determines the virolence of Aggregatibacter actinomycetemcomitans (Aa) via its serotypes and supports the choice of antibiotic preparation.
Bacteria detection with the PadoTest® is essential for:
Periodontitis is an infectious disease of the periodontium that is triggered by bacteria and intensified by immune reactions. The PadoTest® is used to determine the presence of periodontopathogenic germs and classify them into three degrees of severity of periodontitis (type 1 to 3).
This classification allows targeted treatment to be initiated, as the PadoTest® provides information on individual measures, including any antibiotics that may be required.
Examined periodontopathogenic bacteria:
Nomenclature
Aggregatibacter actinomycetemcomitans (Aa)
(formerly Actinobacillus actinomycetemcomitans)
Characteristics
Gram-negative, facultative anaerobe, 7 serotypes with different leukotoxin expression (serotype b with high incidence in localized aggressive periodontitis).
Pathogenicity
Highly pathogenic
Virulence factors
Leukotoxin A
Chemotaxis inhibitory factor
Fibroblast inhibition
Bone resorbing toxin
Collagenase
Lipopolysaccharide endotoxin
Occurrence
In case of localized aggressive and chronic periodontitis. The virulence factors promote colonization and establishment of the germ in the oro-pharyngeal area.
Treatment
Antibiotic of choice is amoxicillin. Cannot be definitively eliminated with scaling/root planing.
References
Aberg, C.H., Kelk, P. and Johannsson, A., 2015. Aggregatibacter actinomycetemcomitans: virulence of its leukotoxin and association with aggressive periodontitis. Virulence, 6 (3), 188-195.
Maeda, H., Fujimoto, C., Haruki, Y., Maeda, T., Kokeguchi, S., Petelin, M., Arai, H., Tanimoto, I., Nishimura, F. and Takashiba, S., 2003. Quantitative real-time PCR using TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, tetQ gene and total bacteria. FEMMS Immunol Med Microbiol., 39 (1), 81-86.
Malik, R., Changela, R., Krishan, P., Gugnani, S. and Bali, D., 2015. Virulence factors of Aggregatibacter actinomycetemcomitans – A status update. Journal of ICDRO, 7 (2), 137-145.
Nomenclature
Aggregatibacter actinomycetemcomitans (Aa) - serotypes a -f
Characteristics
6 serotypes with different leukotoxin expression (serotype b with high incidence in localized aggressive periodontitis).
Pathogenicity
non-virulent (serotype a) to highly virulent (serotype b with JP2)
Virulence factors
Leukotoxin expression
Occurrence
In case of localized aggressive and chronic periodontitis. The virulence factors promote colonization and establishment of the germ in the oro-pharyngeal area.
Treatment
Antibiotic of choice is amoxicillin. Cannot be definitively eliminated with scaling/root planing.
References
Aberg, C.H., Kelk, P. and Johannsson, A., 2015. Aggregatibacter actinomycetemcomitans: virulence of its leukotoxin and association with aggressive periodontitis. Virulence, 6 (3), 188-195.
Maeda, H., Fujimoto, C., Haruki, Y., Maeda, T., Kokeguchi, S., Petelin, M., Arai, H., Tanimoto, I., Nishimura, F. and Takashiba, S., 2003. Quantitative real-time PCR using TaqMan and SYBR Green for Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, tetQ gene and total bacteria. FEMMS Immunol Med Microbiol., 39 (1), 81-86.
Malik, R., Changela, R., Krishan, P., Gugnani, S. and Bali, D., 2015. Virulence factors of Aggregatibacter actinomycetemcomitans – A status update. Journal of ICDRO, 7 (2), 137-145.
Nomenclature
Filifactor alocis (Fa)
Characteristics
gram-positive anaerobes, extremely difficult to cultivate
Pathogenicity
Highly pathogenic
Virulence factors
15 different proteases, microbial surface components that recognize the adhesive matrix (MSCRAMMs), F. alocis complement inhibitor (FACIN).
Occurrence
Occurs in subgingival samples in chronic periodontitis (CP) and generalized aggressive periodontitis (GAP) and is associated with supporting tissue loss. Fa is a cause of chronic infection and induces the production of proinflammatory cytokines that lead to apoptosis of gingival epithelial cells. In comparison with the representatives Pg, Td, Tf, described in the literature as "red complex", which are associated to the highest degree with periodontal diseases, Fa revealed the third highest prevalence in a group of CAP patients and the second highest prevalence in that of CP patients. Fa is primarily located in the regions of the biofilm that originated from the middle or even apical part of the pocket (Schlafer, 2014).
Fa is always intimately intertwined with other periodontal bacteria and exhibits a symbiotic relationship with Pg, for example, which can result in a structural increase of the biofilm.
Treatment
Antibiotic of choice: metronidazole
References
Aruni, A.W., Roy, F. and Fletcher, H.M., 2011. Filifactor alocis Has Virulence Attributes That can Enhance Ist Persistence under Oxidative Stress Conditions and Mediate Invasion of Epithelial Cells by Porphyromonas gingivalis. Infection and Immunity, 79 (10), 3872-3886.
Aruni, W., Chioma, O. and Fletcher, H.M., 2014. Filifactor alocis: The Newly Discovered Kid on the Block with Special Talents. Journal of Dental Research, 93 (8), 725-732.
Aruni, W., Mishra, A., Dou, Y., Chioma, O., Hamilton, B.N. and Fletcher, H.M., 2015. Filifactor alocis - a new emerging periodontal pathogen. Microbes and Infection, 17 (7), 517-530.
Jusko, M., Miedzak, B., Ermert,D., Magda, M., King, B.C., Bielecka, E., Riesbeck, K., Eick, S., Potempa, J. and Blom, A.M., 2016. FACIN, a Double-Edged Sword oft he Emerging Periodontal Pathogen Filifactor alocis: A Metabolic Enzyme Moonlighting as a Complement Inhibitor. Journal of Immunology, 197, 3245-3259.
Schlafer, S., 2014. Filifactor alocis - der noch unterschätze Parodontitis-Keim in der Zahnfleischtasche. Zahnmedizin Report, 9:3.
Nomenclature
Porphyromonas gingivalis (Pg)
Characteristics
gram-negative strict anaerobe
Pathogenicity
highly pathogenic
Virulence factors
Membrane-associated proteases that, for example, cleave fibrinogen, leading to bleeding on probing (BOP), release haemin and iron, and at the same time serve as a food source for further proliferation of the germ. Lipopolysaccharides, exopolysaccharides, OMP (Outer membrane Proteins), collagenase, trypsin-like protease, gelatinase, aminopeptidase.
Occurrence
The germ is not part of the "normal" oral flora; the immune system is not able to completely control an infection. It colonizes the periodontium more evenly than A. actinomycemtemcomitans, which tends to occur at individual sites.
Treatment
Can usually be removed with root planing. If it is still detected after this treatment, reintervention with curettage or chriurgy is indicated. If Pg and Aa are present in larger amounts, antibiotics must be prescribed in addition to scaling/root planing. The antibiotics of choice are metronidazole or ornidazole + amoxicillin.
References
How, K.Y., Song, K.P.and Chan, K.G., 2016. Porphyromonas gingivalis: An Overview of Periodontopathic Pathogen below the Gum Line. Frontiers in Microbiology, 7 (53).
Lyons, S.R., Griffen, A.L. and Leys, E.J., 2000. Quantitative Real-Time PCR for Porphyromonas gingivalis and Total Bacteria. Journal of Clinical Microbiology, 38 (6), 2362-2365.
Mysak, J., Podzimek, S., Sommerova, P., Lyuya-Mi, Y., Bartova, J., Janatova, T., Prochazkova, J. and Duskova, Jana, 2014. Porphyromonas gingivalis: Major Periodontopathic Pathogen Overview. Journal of Immunology Research, ID 476068, 8
Nomenclature
Prevotella intermedia (Pi)
Characteristics
gram-negative obligate anaerobes
Pathogenicity
highly pathogenic
Facteurs de virulence
Exopolysaccharides (EPS)
Cysteine protease (Interpain A)
Occurrence
Is detected as a (co-)pathogen of mostly mixed dentoalveolar infections. Is also referred to as an early marker germ, which creates the anaerobic environment necessary for the colonization of the main periodontal germs by metabolizing the residual sugars in the sulcus or in the periodontal pocket that forms. Pi uses steroid hormones as growth factors, so it might be more prevalent among pregnant women.
Treatment
Pi cannot be treated with scaling/root planing if the bacterial count is elevated. Nitromimidazole preparations (e.g. metronidazole) are the antibiotics of choice. Penicillinase-related penicillin resistance is known in sporadic cases; apart from this, anaerobic ß-lactam antibiotics (e.g. penicillin G, ampicillin, amoxicillin) can be effectively used. The efficacy of clindamycin (resistances) and tetracyclines is different; aminoglycosides are always ineffective.
References
Potempa, M., Potempa, J., Kantyka, T., Nguyen, KA., Wawrzonek, K., Manadhar, S.P., Popadiak, K., Riesbeck, K., Eick, S. and Blom, A.M., 2009. Interpain A, a Cysteine Protease from Prevotella intermedia Inhibits Complement by Degrading Complement Factor 3. PLoS Pathogens, 5 (2), e1000316.
Riggio, M.P., Lennon, A. and Roy, K.M., 1998. Detection of Prevotella intermedia in subgingival plaque of adult periodontitis patients by polymerase chain reaction. Journal Periodontal Research, 33 (6), 369-376.
Yamanaka, T., Yamane, K., Furukawa, T., Matsumoto-Mashimo, C., Sigimori, C., Obata, N., Walker, C.B., Leung, K.P. and Fukushima, H., 2011. Comparision oft he virulence of exopolysaccharide- producing Prevotella intermedia to exopolysaccharide non- producing perodontoipathic organism. BMC Infection Disease, 11, 228-237
Nomenclature
Tannerella forsythia (Tf)
Features
anaérobie strict gram-négatif
Pathogenicity
highly pathogenic
Virulence factors
3 proteolytic enzymes: cysteine protease (PrtH), carilysin (structurally comparable to human matrix metalloproteases), mirolase (calcium-dependent serine protease).
Occurrence
Characteristic of severe bone defects. In "active" bags in significantly higher numbers than in "inactive" bags. Also known in cases of recurrent periodontitis. Often associated with refractory periodontal disease. There is a close relationship of co-occurrence with Treponema denticola and Porphyromonas gingivalis.
Treatment
can usually be removed with root planing. The antibiotics of choice are metronidazole or ornidazole, if explicitly necessary (especially in the case of severe inflammatory symptoms).
References
Ksiazek, M., Mizgalska, D., Eick, S., Thorgersen, I.B., Enghild, J.J. and Potempa, J. 2015. KLIKK proteases of Tannerella forsythia: putative virulence factors with a unique domain structure. Frontiers in Microbiology, 6, 312.
Saito, D., Coutinho, L.L., Borges Saito, C.P., Tsai, S.M., Höfling, J.F. and Goncalves, R.B., 2009. Real-time polymerase chain reactionquantification of Porphyromonas gingivalis and Tannerella forsythia in primary endodontic infections. Journal of Endodontics, 35 (11), 1518-1524.
Sharma, A., 2000. Virulence mechansims of Tannerella forsythia. Periodontology, 54 (1), 106-116.
Nomenclature
Treponema denticola (Td)
Characteristics
short, strictly anaerobic gram-negative spirochete
Pathogenicity
highly pathogenic
Virulence factors
tissue-destroying proteases, hyaluronidases, phosphatases and phopholipases. Stimulates formation of IL-1 alpha and TNF- alpha.
Occurrence
is associated with periodontal destruction (advanced periodontitis), necrotizing ulcerative gingivitis/periodontitis.
Treatment
is useful as a marker in the evaluation of treatment success (scaling/root planing) in therapy-refractory pockets. As antibiotic of choice, if required, metronidazole or ornidazole.
References
Asai, Y., Jinno, T., Igarashi, H., Ohyama, Y. and Ogawa, T., 2002. Detection and Quantification of Oral Treponemes in Subgingival Plaque by Real-Time PCR. Journal of Clinical Microbiology, 40 (9), 3334-3340.
Dashper, S.G., Seers, C.A., Tan, K.H. and Reynolds, E.C., 2011. Virulence Factors oft he Oral Spirochete Treponema denticola. Journal Dent. Research, 90 (6), 691-703.
Pandit, N., Gugnani, S., Sushil, D. and Bali, D., 2016. Treponema denticola: A teammate in periodontal progression. Journal of ICDRO, 8 (1), 58-62.
Sela, M.N., 2001. Role of Treponema denticola in periodontal diseases. Crit Rev Oral Biol Med, 12 (5), 399-413.
For detection and quantification, the PadoTest® uses a special multiplex real-time PCR (polymerase chain reaction). The high precision of this form of analysis provides an image of the examined key germs in the gingival pocket. In terms of price, the PadoTest® is extremely attractive and can be used very flexibly due to the possibility of multi-site or single-site tests.
The practitioner recognizes the damage pattern, the PadoTest® determines the pathogenicity of the bacterial load and recommends the therapeutic measures!