PadoTest® is a microbiological laboratory analysis for dental practices. The procedure determines the bacterial load within the periodontal pocket and identifies the serotypes of Aggregatibacter actinomycetemcomitans (PadoSero). The analytical report provides the practice with quantitative laboratory measurements.
The analysis provides quantitative data on the bacterial profile. These objective measurements serve the practitioner as an informational reference to support individualized treatment planning.
Quantification of marker bacteria enables a comparative evaluation of the microbial status before and after instrumentation, as well as continuous laboratory documentation during supportive periodontal therapy (SPT).
Every PadoTest® analysis routinely includes serotyping of Aggregatibacter actinomycetemcomitans (Aa). The laboratory provides practitioners with differentiated microbiological data at the sub-species level.
In cases of refractory disease courses, the microbiological profile offers additional laboratory reference points to assist in dental diagnostic evaluation.
Periodontitis is a bacteria-induced infection of the periodontium. PadoTest® detects and quantifies specific periodontal marker bacteria. The laboratory report classifies the measured bacterial load into quantitative levels, providing the dental practice with an objective assessment of the microbial colonization status.
Evaluated Marker 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.
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.
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.
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.
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.
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.
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.
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, PadoTest® utilizes a specialized real-time PCR (polymerase chain reaction) method. The high analytical accuracy of this assay provides a precise profile of the evaluated marker bacteria within the periodontal pocket. PadoTest® offers a cost-effective diagnostic solution and flexible application via single-site or pooled multi-site sampling options.
PadoTest® determines the bacterial load, providing objective microbiological findings to support the practitioner in determining the appropriate therapeutic measures.