Identification of
            <i>Actinobacillus actinomycetemcomitans</i>
            Serotypes by Multiplex PCR

Suzuki, Nao; Nakano, Yoshio; Yamaguchi, Yasuo; Ikeda, Daisuke; Kinoshita, Toshihiko

doi:10.1128/jcm.39.5.2002-2005.2001

Cited by 77 publications

(72 citation statements)

References 31 publications

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“…Of the nucleic acid amplification methods, PCR-based amplification methods are widely used for the diagnosis of various diseases. Periodontitis is a common infectious disease, and a PCR-based diagnosis system for periodontitis has been developed (4,42,45). Despite their simplicity and accuracy, PCRbased diagnosis methods are not widely used in private clinics as routine diagnostic tools, due to the need for a thermal cycler.…”

Section: Discussionmentioning

confidence: 99%

“…The cells were resuspended in 100 l of cell lysis buffer and boiled at 100°C for 5 min, and the supernatant served as the template (42).…”

Section: Vol 43 2005 Lamp For Periodontal Pathogens 2419mentioning

confidence: 99%

“…Of the various methods used to diagnose infectious disease (3,6,19,37), molecularbased methods are often used (13,44); of these, the PCR is one of the most widely used techniques (4,42,45). PCR-based detection of bacteria is sensitive and specific, and some PCRderived methods, such as nested PCR (7,9) and PCR-restriction fragment length polymorphism (36), have been developed.…”

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confidence: 99%

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Loop-Mediated Isothermal Amplification Method for Rapid Detection of the Periodontopathic BacteriaPorphyromonas gingivalis,Tannerella forsythia, andTreponema denticola

et al. 2005

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Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method, was developed for the rapid detection of the major periodontal pathogens Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. The LAMP method amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specially designed primers and a DNA polymerase with strand displacement activity. In this study, we initially designed the primers for LAMP assays to detect these bacteria and evaluated the specificity and sensitivity of these assays. The specificities of the primers for these bacteria were examined using various oral bacteria and various reaction times. The lower detection limits of the 60-min LAMP reaction without loop primers were 1 g/tube for P. gingivalis, 10 fg/tube for T. forsythia, and 1 ng/tube for T. denticola. Addition of the loop primers for each bacterium improved the detection specificities and sensitivities by several magnitudes. Furthermore, LAMP assays were applied to the rapid detection of these periodontal pathogens in clinical specimens, and the results were compared with those of conventional PCR detection. The results of the LAMP assays corresponded to those of conventional PCR assays. These results indicate that the LAMP assay is an extremely rapid, highly sensitive, specific method. This method is very useful for the rapid detection of periodontopathic bacteria and the diagnosis of periodontal disease.

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Section: Discussionmentioning

confidence: 99%

“…The cells were resuspended in 100 l of cell lysis buffer and boiled at 100°C for 5 min, and the supernatant served as the template (42).…”

Section: Vol 43 2005 Lamp For Periodontal Pathogens 2419mentioning

confidence: 99%

mentioning

confidence: 99%

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Loop-Mediated Isothermal Amplification Method for Rapid Detection of the Periodontopathic BacteriaPorphyromonas gingivalis,Tannerella forsythia, andTreponema denticola

et al. 2005

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“…Actinobacillus actinomycetemcomitans, a small, nonmotile, facultatively anaerobic coccobacillus, is a major pathogen in periodontitis (1). The species is divided into five genetically diverse (3,27) serotypes (29,23). However, up to every 10th bacterium-positive individual carries a nonserotypeable strain (20).…”

mentioning

confidence: 99%

Antigenically Diverse Reference Strains and Autologous Strains ofActinobacillus actinomycetemcomitansAre Equally Efficient Antigens in Enzyme-Linked Immunosorbent Assay Analysis

et al. 2002

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Actinobacillus actinomycetemcomitans is a major pathogen in periodontitis. Data on the clinical relevance of serum immunoglobulin G (IgG) antibody levels against this species are controversial. The aim of the present study was to elucidate how different strains used as antigens in enzyme-linked immunosorbent assay (ELISA) influence the detection of individuals with elevated serum IgG levels against A. actinomycetemcomitans. We hypothesized that the highest antibody levels are targeted to the autologous strains. A total of 19 strains-six antigenically diverse reference strains (serotypes a through e and a nonserotypeable strain) and 13 serotyped autologous strains-were used as whole-cell antigens in ELISA. Serum samples were from 26 untreated adult patients with periodontitis, whose subgingival bacterial samples were either culture positive (n ‫؍‬ 13) or culture negative (n ‫؍‬ 13) for A. actinomycetemcomitans, and from 10 culture-negative nonperiodontitis subjects. The highest individual (P < 0.05) IgG levels against the reference strains were most commonly against serotypes a and b in patients and against serotype c in nonperiodontitis subjects. The culture-positive patients had the highest (P < 0.05) IgG antibody levels against their autologous strains and against the reference strains of the same serotype. On the contrary, for these patients the levels of antibody against the reference strains of other serotypes were comparable to those of the nonperiodontitis subjects. The results indicated that the serum IgG antibody levels against A. actinomycetemcomitans strongly depend on the strains used as antigens in the ELISA. Elevated serum IgG levels against A. actinomycetemcomitans can be detected equally well using either the autologous strains or a variety of antigenically diverse reference strains as antigens.

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“…En este contexto, la mayor frecuencia de detección del serotipo b de A. actinomycetemcomitans en pacientes con periodontitis agresiva sugiere un mayor potencial patogénico y este podría asociarse a una mayor inmunogenicidad y virulencia (33) . En efecto, el serotipo b evade más eficazmente la fagocitosis por macrófagos (34) , inhibe con mayor intensidad la proliferación de macrófagos y neutrófilos (35) , produce mayores niveles de microvesículas citotóxicas (36) e induce un incremento en la secreción de IgG, IgM e IgA por linfocitos B activados (29) . En efecto, el LPS es importante en A. actinomycetemcomitans dado que induce varibailidad en la respuesta inmune desplegada por el hospedero, a diferencia de los demás factores de virulencia (leucotoxina, toxina distensora citoletal, entre otros).…”

Section: Introductionunclassified

Variabilidad de la síntesis de citoquinas por células dendríticas humanas estimuladas con los distintos serotipos de Aggregatibacter actinomycetecomitans

Díaz‐Zúñiga

Melgar‐Rodríguez

et al. 2013

Rev. Clin. Periodoncia Implantol. Rehabil. Oral

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Identification of Actinobacillus actinomycetemcomitans Serotypes by Multiplex PCR

Cited by 77 publications

References 31 publications

Loop-Mediated Isothermal Amplification Method for Rapid Detection of the Periodontopathic BacteriaPorphyromonas gingivalis,Tannerella forsythia, andTreponema denticola

Loop-Mediated Isothermal Amplification Method for Rapid Detection of the Periodontopathic BacteriaPorphyromonas gingivalis,Tannerella forsythia, andTreponema denticola

Antigenically Diverse Reference Strains and Autologous Strains ofActinobacillus actinomycetemcomitansAre Equally Efficient Antigens in Enzyme-Linked Immunosorbent Assay Analysis

Variabilidad de la síntesis de citoquinas por células dendríticas humanas estimuladas con los distintos serotipos de Aggregatibacter actinomycetecomitans

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