Recovery of A. actinomycetemcomitans from teeth, tongue, and saliva

The recovery of actinobacillus actinomycetemcomitans simultaneously from subgingival sites around teeth and dorsum of the tongue and/or saliva was examined in 293 subjects at 444 visits; 295 paired samples were available from subgingival sites and tongue, 171 paired samples from subgingival sites and stimulated saliva, and 137 paired samples from subgingival sites and unstimulated saliva. Sixty-one subjects were periodontally healthy (mean age 20.3 years); 55 exhibited localized juvenile periodontitis (mean age 21.8 years); 176 adult periodontitis (mean age 46.7 years); and 1 prepubertal periodontitis (age 10 years). When A. actinomycetemcomitans was recovered from subgingival sites, it was also found in 56.3%, 69.9%, and 35.9% of the paired samples from tongue, and stimulated and unstimulated saliva, respectively. No difference in the detection rate of A. actinomycetemcomitans from tongue or stimulated saliva was seen between the subjects with healthy or diseased periodontium. When A. actinomycetemcomitans was not recovered from subgingival sites, it was cultured in 6.8%, 2.0%, and 1.4% of the paired samples from tongue, and stimulated and unstimulated saliva, respectively. In search for noninvasive, inexpensive, and easily run sampling methods for the recovery of oral A. actinomycetemcomitans samples from stimulated saliva and tongue may prove useful in clinical periodontology.     

Sampling strategy for intraoral detection of periodontal pathogens before and following periodontal therapy

BACKGROUND: The aim of this study was to identify a sampling strategy with high probability for detecting oral colonization by Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Eikenella corrodens, Tannerella forsythensis, Prevotella intermedia, Prevotella nigrescens, and Treponema denticola before and following mechanical periodontal therapy. METHODS: Samples were taken from the following intraoral sites in 35 patients with untreated chronic periodontitis before and 1.5, 3, and 6 months after non-surgical periodontal therapy: supra- and subgingival plaque from the deepest pockets in each sextant; pooled supra- and subgingival plaque from another six randomly selected, less affected teeth; mucosal swab samples from the tongue, tonsils, throat, and buccal mucosa; and stimulated and unstimulated saliva. Microbial species were identified by polymerase chain reaction (PCR). RESULTS: Of the sampling of all assessed sites, the highest probability for simultaneously detecting the tested pathogens was found in respect to the combination of supra- and subgingival plaque samples taken from the most affected tooth in each sextant in untreated patients (probability: 83% to 95% for the assessed bacteria). These results were consistently observed throughout the study period. CONCLUSIONS: For determining the intraoral carrier state of patients with periodontitis, a combined sample of supra- and subgingival plaque taken from the deepest periodontal pocket in each sextant may yield the most reliable result. This sampling strategy may be used in routine microbial testing and clinical research.     

Detection of Epstein-Barr virus and human cytomegalovirus in blood and oral samples: comparison of three sampling methods

The purpose of this study was to identify and compare the presence of HCMV and EBV-1 in subgingival plaque, unstimulated saliva and peripheral blood of patients with chronic periodontitis. Forty patients diagnosed with chronic periodontitis (mean age, 41.7 years) were recruited. Unstimulated saliva, subgingival plaque and peripheral blood were collected from each patient and the DNA of each sample was isolated. The viruses were detected using the nested PCR technique. The detection frequency of EBV-1 in subgingival plaque, saliva and peripheral blood was 45%, 37.5% and 25%, respectively. HCMV was detected in 82.5% of subgingival plaque samples and peripheral blood and in 75% of salivary samples. The sensitivity for detecting EBV-1 in saliva and peripheral blood when EBV-1 was detected in subgingival plaque samples was low (22% and 27.7%, respectively) and the sensitivity for detecting HCMV in saliva and peripheral blood when compared to subgingival plaque was high (81.8% and 87.8%, respectively). There is a high agreement among the three sampling methods in detection of HCMV, but the detection of EBV-1 would require a combination of saliva and subgingival plaque sampling to avoid false negative results.     

Actinobacillus actinomycetemcomitans recovery from extracrevicular locations of the mouth

Associations between recovery of Actinobacillus actinomycetemcomitans from samples of subgingival plaque, and samples of buccal mucosa, tongue and unstimulated saliva were studied in 107 subjects. Ten subjects had gingivitis, 18 localized juvenile periodontitis, 45 rapidly progressive periodontitis and 32 adult periodontitis. Two children suffered from prepubertal periodontitis. Heterogeneity tests for associations in different study populations yielded nonsignificant results. Mantel-Haenszel's common odds ratios were 52.9, 37.2 and 19.8 for respective associations between pooled subgingival samples, and cheek, saliva and tongue samples. Significant McNemar's chi-square of 5.88, 11.25 and 16.96 for respective associations pointed to secondary occurrence of A. actinomycetemcomitans in extra-crevicular samples. Multiple linear regression yielded a significant influence of the number of deep periodontal pockets of 7 mm or more and a negative influence of the diagnosis "adult periodontitis" on the log-transformed number of colony-forming units of A. actinomycetemcomitans in samples from cheek mucosa in patients infected with the organism. Extracrevicular occurrence of A. actinomycetemcomitans seems to reflect total subgingival numbers of the organism. Especially sampling check mucosa appears to be a promising tool in the diagnosis of a periodontal infection with A. actinomycetemcomitans .     

Aggregatibacter actinomycetemcomitans as indicator for aggressive periodontitis by two analysing strategies

OBJECTIVE: To compare the subgingival microbiota of aggressive and chronic periodontitis (ChP) using single-site and pooled plaque samples. METHODS: In 60 patients with aggressive or ChP, subgingival plaque was sampled from the four deepest pockets using two sterile paper points simultaneously. One paper point from each pocket was put in a separate transport vial, the second was pooled with the three other paper points of a respective patient. The content of each vial was analysed for Aggregatibacter actinomycetemcomitans, Tannerella forsythensis, Porphyromonas gingivalis, and Treponema denticola. RESULTS: Pooled plaque samples detected higher numbers for all tested pathogens than single-site samples. Detection frequencies were similar for both strategies. Using single-site samples, A. actinomycetemcomitans detection rate was statistically significantly a higher in aggressive than in ChP (p=0.01). A. actinomycetemcomitans was found in higher numbers, the other pathogens in lower numbers in aggressive than in ChP. Neither presence nor absence of one of the tested bacteria had sufficient positive or negative predictive value for aggressive periodontitis. CONCLUSION: A. actinomycetemcomitans was detected in higher numbers and frequency in aggressive than in ChP. Its detection may confirm the clinical diagnosis and influence therapy. As a diagnostic test, its sensitivity and predictive value was low.     

Clinical responses to mechanical periodontal treatment in Chinese chronic periodontitis patients with and without Actinobacillus actinomycetemcomitans

BACKGROUND: The purpose of this study was to compare 12-month clinical responses to mechanical periodontal treatment in Chinese chronic periodontitis patients at sites with and without Actinobacillus actinomycetemcomitans at baseline, and to investigate the ability of mechanical periodontal treatment to eliminate A. actinomycetemcomitans. METHODS: Nineteen patients and a total of 76 selected sites with a mean probing depth (PD) of > or = 7 mm were studied. Whole mouth presence or absence of supragingival plaque (PI%), bleeding on probing (BOP%), probing depth (PD), and probing attachment level (PAL) were recorded at six sites per tooth at baseline and after 3, 9, and 12 months. Baseline subgingival plaque samples were taken from the deepest PD site in each quadrant using sterile paper points and were cultured on TSBV plates for 5 days in a 5% CO2-air incubator. All sites received mechanical periodontal treatment, which included oral hygiene instructions and supragingival and subgingival instrumentation with or without surgical access, with maintenance care being provided once every 3 months thereafter. RESULTS: At baseline, A. actinomycetemcomitans was isolated in 13 of the 19 subjects (68%) and in 29 out of the 76 sampled sites (38%). At the end of 12 months, in three of the initially A. actinomycetemcomitans-positive subjects, A. actinomycetemcomitans was not detected in the sampled sites, while one subject, in whom A. actinomycetemcomitans was not initially found at the sampled sites was A. actinomycetemcomitans-positive at 12 months. Multi-level variance component models showed there was no statistically significant difference in all clinical parameters between A. actinomycetemcomitans-positive and -negative subjects (P > 0.05). In the sampled sites of the initially A. actinomycetemcomitans-positive subjects, the mean PD was reduced from 7.6 +/- 1.6 mm to 3.2 +/- 1.8 mm, the mean PAL gain was 1.4 +/- 2.0 mm, and the mean recession was 3.0 +/- 2.3 mm. The corresponding figures in the sampled sites of the initially A. actinomycetemcomitans-negative subjects were 7.5 +/- 1.6 mm to 2.7 +/- 1.0 mm, 2.3 +/- 2.6 mm and 2.4 +/- 2.2 mm for mean PD changes, PAL gain, and mean recession, respectively. CONCLUSIONS: Favorable clinical responses to mechanical periodontal therapy may occur in Chinese chronic periodontitis patients at sites infected with A. actinomycetemcomitans. The mere detection of subgignival A. actinomycetemcomitans does not necessarily imply poorer treatment outcomes in the control of chronic periodontitis.     

Quantitative analysis of periodontal pathogens in aggressive periodontitis patients in a Japanese population

The aim of the present study was to clarify the relationship between the relative/absolute numbers of periodontal bacteria and different types of periodontitis. Fifteen patients with localized aggressive periodontitis (LAgP), 25 patients with generalized aggressive periodontitis (GAgP) and 28 patients with chronic periodontitis (CP) were included in this study. Saliva and subgingival plaque samples were collected from all subjects for microbiological analysis. The prevalence and proportions of Actinobacillus actinomycetemcomitans, Tannerella forsythensis, Porphyromonas gingivalis and Treponema denticola were determined by conventional PCR and real-time PCR. The prevalence of A. actinomycetemcomitans in saliva was significantly higher in LAgP patients (46.7%) and GAgP patients (40.0%) than that in CP patients (14.3%). The mean proportion of A. actinomycetemcomitans in LAgP patients (4.42%) was significantly higher than that in GAgP patients (0.59%) and CP patients (0.37%) in saliva. In subgingival plaque, LAgP patients showed a significantly higher mean proportion of T. forsythensis (19.8%) than CP patients (7.45%). In conclusion, A. actinomycetemcomitans was the more predominant periodontopathic bacteria in LAgP than in GAgP and CP. The increased proportion of T. forsythensis might relate to LAgP, in addition to A. actinomycetemcomitans. These results indicate that real-time PCR analysis is useful for the evaluation of the bacterial profiles in different types of periodontitis.     

PCR方法对牙周炎患者唾液中牙龈卟啉单胞菌的检测

目的用PCR方法,检测牙周病患者唾液中牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g),并探讨采用唾液标本与龈下菌斑标本检测结果的一致性.方法选择临床54例牙周炎患者病例,分别取其静止唾液和龈下菌斑标本,设计P.g菌16SrDNA引物,分别对2种标本进行PCR扩增,观察P. g菌的检出率,并计算kappa值.结果静止唾液和龈下菌斑标本中P.g菌的检测结果具有高度一致性,其检出率分别为83.3%(45/54)和79.6%(43/54),Kappa值为0.755,准确度达92.6%.结论本研究所用的引物可用于口腔中P.g菌的检测,特别是研究中采用的唾液标本,取材方便,有可能代替龈下菌斑标本.     

Cytolethal distending toxin of Actinobacillus actinomycetemcomitans. Occurrence and association with periodontal disease

The cytolethal distending toxin (Cdt) of Actinobacillus actinomycetemcomitans, a periodontal pathogen, is a newly described cytotoxin with immunosuppressive properties, capable of causing cell cycle arrest of lymphocytes. The objectives of this study were to investigate the occurrence of A. actinomycetemcomitans with the cdt genotype in the subgingival plaque of periodontitis patients and to determine the association of this bacterial genotype with periodontal disease. A total of 146 subgingival plaque samples from periodontitis patients were assayed by the PCR method using oligonucleotide primers targeting the cdt operon of A. actinomycetemcomitans. Primers targeting the leukotoxin gene A (ltxA) of A. actinomycetemcomitans was used to determine the occurrence of the bacteria in the plaque samples at baseline. At baseline, A. actinomycetemcomitans was detected in 106 out of 146 (73%) diseased sites studied. Among the 106 diseased sites found to harbor A. actinomycetemcomitans, 13 sites were positive for the bacteria with the cdt genotype (12%). Out of the 13 positive sites, 10 sites were obtained from patients diagnosed with aggressive periodontitis (77%). Thus, A. actinomycetemcomitans with the cdt genetic subtype has low occurrence in the subgingival plaque of periodontitis patients. However, a strong association was observed between the presence of the bacteria and aggressive forms of periodontitis. Thus, the cytotoxic and immunosuppressive properties of A. actinomycetemcomitans Cdt may function to cripple the host immunity and contribute to the pathogenesis of aggressive periodontitis.     

Comparison of cultural methods and DNA probe analyses for the detection of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius in subgingival plaque samples

The purpose of this study was to compare DNA probe analyses to cultural methods for detecting three periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius, in human subgingival plaque. Subgingival sites from patients diagnosed as either healthy or showing evidence of gingivitis or juvenile or adult periodontitis were sampled using two paper points. The number of these pathogens from one paper point was determined using microbiologic media and speciated by biochemical tests. Results were then compared to bacterial numbers obtained from the other paper point using species-specific DNA probes. In 60 samples from the disease group, DNA probe analysis demonstrated 100% effectiveness in detecting A. actinomycetemcomitans and B. intermedius and 91% effectiveness in detecting B. gingivalis at culture positive levels (greater than or equal to 10(3) cells). In addition, probe assays frequently identified these pathogens in samples that were culture negative. Probe analysis revealed a better correlation between presence of a pathogen and clinical evidence of disease on an individual patient basis. In contrast, most samples taken from sites of healthy individuals showed undetectable levels of all three pathogens as determined by both techniques. These results suggest that DNA probe technology is at least equivalent and often superior to cultural methods for detecting A. actinomycetemcomitans, B. gingivalis, and B. intermedius in human subgingival plaque samples.     

Cytopathic effects of Actinobacillus actinomycetemcomitans on monkey blood leukocytes

The suitability of cynomolgus monkeys (Macaca fascicularis) for studies concerned with the biologic properties of Actinobacillus actinomycetemcomitans is the subject of the present investigation. We found that normal monkeys harbored leukotoxic strains of A. actinomycetemcomitans in subgingival plaque samples. Monkey peripheral blood PMNs and monocytes were killed following in vitro exposure to sonic extracts of leukotoxic strains of A. actinomycetemcomitans . Monkey sera were capable of inhibiting the leukotoxic properties of A. actinomycetemcomitans sonic extracts due to the presence of IgG antibodies which neutralized the leukotoxin. Similarly, sera from patients with juvenile periodontitis (but not normal human sera) abolished leukotoxin-mediated killing of monkey PMNs. Monkey peripheral blood lymphoid cells were not killed by A. actinomycetemcomitans but demonstrated depressed responses to mitogens following pre-incubation with A. actinomycetemcomitans sonicates prepared from either leukotoxic or "non-leukotoxic" human strains. These studies suggest that cynomolgus monkeys may serve as a suitable in vitro and in vivo model for delineating more about host- A. actinomycetemcomitans interrelationships in the etiology of human periodontal disease.     

Prevalence of Actinobacillus actinomycetemcomitans in an ethnic adult Chinese population

AIM: The aim of this study was to determine the prevalence and the structure of the leukotoxin promoter region of Actinobacillus actinomycetemcomitans in an ethnic Chinese population. METHOD: Subgingival plaque samples were collected from 42 patients with moderate to advanced periodontitis and 50 periodontally healthy patients. A. actinomycetemcomitans was detected directly from the crude subgingival plaque by PCR using leukotoxin gene specific primers. The presence of A. actinomycetemcomitans was determined by a single 285 bp PCR amplicon. RESULTS: A. actinomycetemcomitans was found to be present in the subgingival plaque of 68 out of a total of 92 patients examined (74%). 29 out of the 42 periodontitis patients tested were carriers of A. actinomycetemcomitans (69%). Among the periodontally healthy patients studied, 39 out of 50 subjects possessed the bacteria (78%). PCR analysis of the promoter region of the ltx operon revealed that none of the 42 moderate to advanced periodontitis patients examined harboured A. actinomycetemcomitans strains with the JP2-like promoter of the ltx operon, known to enhance leukotoxin expression. 2 out of the 27 advanced periodontitis patients clinically diagnosed as suffering from rapidly progressive periodontitis were found to be carriers of the mildly toxic strain of A. actinomycetemcomitans with the characteristic 652-like promoter. CONCLUSIONS: The high prevalence of A. actinomycetemcomitans, regardless of whether the subgingival samples were analysed from patients with healthy or diseased periodontium suggests that this bacterial species is part of the normal oral flora of ethnic Chinese. Our preliminary results also suggested that subjects who harboured the mildly toxic strain of A. actinomycetemcomitans were potentially susceptible to aggressive forms of periodontitis.     

伴放线放线杆菌在龈下菌斑和颊黏膜分布研究

目的:比较伴放线放线杆菌(actinobac illus actinomycetem com itans,A.a)在不同类型牙周炎患者龈下菌斑和颊黏膜中的分布。方法:通过聚合酶链反应(polym erase chain reaction,PCR)对侵袭性牙周炎患者(AgP)、慢性牙周炎患者(CP)、牙周健康者口腔龈下菌斑和颊黏膜中的A.a进行检测,分析该菌分别在两部位的相对含量。结果:AgP组菌斑和颊黏膜样本中A.a阳性检出率均为41.7%,分别高于CP组(菌斑16.7%、颊黏膜10.0%)和牙周健康组(菌斑和颊黏膜均为0%)。AgP组A.a在菌斑和颊黏膜的相对含量分别为38.5%和22.2%,高于CP组(菌斑19%、颊黏膜12.75%)。结论:A.a不仅存在于龈下菌斑中,也能够粘附于颊黏膜;A.a是AgP的主要优势菌也参与了CP的菌群组成。     

Morphological compositions of subgingival microbiota in Actinobacillus actinomycetemcomitans- associated periodontitis

Infrequent occurrence of spirochetes and rather low proportions of these organisms have been reported in localized juvenile periodontitis, where periodontal lesions often harbour large numbers of Actinobacillus actinomycetemcomitans, a suspected principal periopathogen strongly implicated in the pathogenesis of this and other forms of chronic periodontitis. We studied the association of subgingival A. actinomycetemcomitans with the morphological composition of the subgingival microbiota in a large population of patients suffering from advanced periodontitis. Subgingival plaque from the deepest pockets of every quadrant of their dentitions was sampled and pooled in 70 patients between 14 and 63 years of age, and analysed morphologically by phase-contrast microscopy. A minimum % similarity index was employed to define 4 clusters with different morphological composition of the floras. The actual proportion of A. actinomycetemcomitans was determined at 2 sites with deep periodontal pockets. All clusters harboured patients infected with A. actinomycetemcomitans. If present, in clusters predominated by motile rods or medium-sized spirochetes, the organism was found in rather low proportions (median 5.3% and 3.4%, respectively). However, the cluster with a pooled flora mainly consisting of coccoid cells revealed periodontal sites with A. actinomycetemcomitans in proportions of more than 53% (median), if the organism was present (p less than 0.01). We found a positive correlation between proportions of A. actinomycetemcomitans and cocci (R = 0.65) and negative correlations with spirochetes and motile rods (R = 0.61, R = -0.59, respectively). Cautious interpretation of subgingival plaque predominated by coccoid cells is recommended, if deep periodontal pockets and obvious signs of inflammation are present, since these pockets were found to be often infected with large numbers of A. actinomycetemcomitans.     

伴放线菌嗜血菌在慢性牙周炎患者和牙周健康者中的分布

目的分析伴放线菌嗜血菌在慢性牙周炎患者和牙周健康者中的分布情况。方法选择116例慢性牙周炎患者（CP组）和111例牙周健康者（健康组）为研究对象。CP组选取磨牙区牙周袋最深的2个患牙的探诊深度最深点作为患牙的取样位点,同时选取磨牙或前磨牙区1个健康牙的近中位点作为健康牙的取样位点;健康组选取上颌第一磨牙近中位点作为取样位点。收集取样位点的龈下菌斑,提取DNA,用16S rRNA聚合酶链反应检测伴放线菌嗜血菌的分布;同时检查并记录取样牙的牙周临床指数（包括探诊深度、临床附着丧失和探诊出血）,分析伴放线菌嗜血菌检出率和牙周临床指数的关系。结果CP组患牙伴放线菌嗜血菌检出率为33.62%,明显高于CP组健康牙和健康组（P<0.01）。伴放线菌嗜血菌检出率随着患者年龄的增加而降低,随着探诊深度、临床附着丧失的增加而增加,探诊出血阳性患牙的检出率（37.07%）也明显高于探诊出血阴性的患牙（7.41%）（P<0.05）。结论伴放线菌嗜血菌检出率随着牙周炎症程度的加重而升高,与慢性牙周炎的发生发展具有密切的联系。     

牙周炎患者唾液中伴放线放线杆菌的检出状况分析

目的 检测不同类型牙周炎患者唾液中的伴放线放线杆菌(Actinobacillusactinomycetemcomitans,Aa),探讨唾液和集合龈下菌斑中Aa检出率的差异以及唾液中Aa的存在状况与牙周临床指标的关系. 方法 收集50例侵袭性牙周炎(aggressive periodontitis,AgP)患者、48例慢性牙周炎(chronic periedontitis,CP)患者和25例非牙周炎者的非刺激性全唾液和集合龈下菌斑,应用聚合酶链反应(PcR)技术检测两种样本中的Aa. 结果 Aa在AgP患者唾液中的检出率(32%)显著高于非牙周炎者(4%)和CP患者(15%),差异均有统计学意义(P<0.01,P<0.05),同时Aa在AgP患者唾液中的检出率也显著高于集合龈下菌斑样本(16%),差异亦有统计学意义(P<0.05).年龄≤30岁是唾液中存在Aa的危险指征(OR=3.23,P<0.05);出血指数≥3的位点超过70%与唾液中存在Aa有关(OR=19.21,P<0.01). 结论 AgP患者唾液样本中Aa的检出率明显高于集合龈下菌斑样本,亦高于CP患者和非牙周炎者,提示Aa可能参与AgP的发生和发展.     

Characterization of Actinobacillus actinomycetemcomitans isolated from young Chinese aggressive periodontitis patients

OBJECTIVE: This study characterized Actinobacillus actinomycetemcomitans isolates from young Chinese aggressive periodontitis patients. METHODS: Subgingival plaque samples (two/subject) were collected from diseased subjects < 25 years old (n = 9, mean age 21.1 +/- 1.6 years) and age-matched periodontitis-free controls (n = 47, mean age 22.0 +/- 1.1 years). Selective and anaerobic culture were used. The serotype, leukotoxin gene (ltx) operon promoter and the cytolethal distending toxin (cdt) genes complex of the A. actinomycetemcomitans isolates were investigated. Effects of the isolates on non-keratinizing periodontal ligament epithelial cells monolayer were studied. RESULTS: Diseased subjects had significantly higher full-mouth bleeding score (p = 0.002) and total viable counts from plaque samples (7.2 x 10(6) vs. 2.1 x 10(5) CFU/paperpoint, p < 0.005). A. actinomycetemcomitans was isolated from 67%/56% or 6%/4% of diseased or controls subject/sites, respectively (p < 0.001). The proportion of A. actinomycetemcomitans isolatable from aggressive periodontitis or periodontitis-free associated subgingival plaque was low (0.7% vs. 0.1%, p < 0.02). The serotype of the isolates was characterized. All isolates possessed 652-like ltx gene promoter and all but one serotype c isolate from a diseased patient had intact cdtABC genes. That particular strain appeared to confer the least cellular damages on periodontal ligament epithelial monolayer compared to others. CONCLUSION: This preliminary study confirmed the notion of increased prevalence and quantity of A. actinomycetemcomitans associated with aggressive periodontitis in young patients. The overall ltx promoter and cdt characteristics of the A. actinomycetemcomitans isolates, however, were similar among the diseased and control groups. A strain lacking the cdtABC gene appeared to be less damaging to a periodontal ligament epithelial cell model. Further studies therefore are warranted to clarify the pathogenic role and potentials of A. actinomycetemcomitans in aggressive periodontitis.     

Periodontitis lesions are a source of salivary cytomegalovirus and Epstein-Barr virus

AIM: Several herpesvirus species can be detected in periodontal pockets and saliva. This study compared human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) DNA copy counts in periodontitis sites and in whole saliva, and evaluated the potential of periodontal therapy to reduce the salivary level of the two viruses. MATERIAL AND METHODS: A total of 20 systemically healthy periodontitis patients, 21-56 years of age, participated in the study. All 20 patients were examined at baseline, and seven patients also at 3 months after periodontal therapy. Treatment included oral hygiene instruction, scaling and root planing, and surgery. Clinical parameters were evaluated using established methods. In each patient, virological samples were collected from one periodontal pocket of 6-10 mm probing depth, from the adjacent inflamed periodontal pocket wall, and from unstimulated whole saliva. Relationships between subgingival, gingival tissue and salivary herpesvirus counts were evaluated using Spearman's and Kendall's rank correlation coefficient tests. The 5'-nuclease (TaqMan) real-time polymerase chain reaction (PCR) assay was employed to quantify genomic copies of periodontal HCMV and EBV. RESULTS: At baseline, the 20 periodontitis patients showed significant positive correlations between gingival tissue and salivary counts of HCMV DNA (p=0.003) and EBV DNA (p=0.045). Periodontal pocket depth was positively correlated with salivary EBV DNA counts (p=0.002). Periodontal therapy reduced average full-mouth periodontal pocket depth from 4.6 mm to 1.4 mm, plaque index from 2.1 to 0.9, and gingival index from 2.1 to 0.4. Following treatment, HCMV DNA counts decreased 37.5 fold in subgingival sites and 64.6 fold in saliva, and EBV DNA counts decreased 5.7 fold in subgingival sites and 12.9 fold in saliva. CONCLUSIONS: The present study provides compelling evidence of a periodontitis source for salivary HCMV and EBV. The potential of periodontal therapy to decrease herpesvirus salivary counts may help diminish herpesvirus transmission from person to person and herpesvirus-related diseases in exposed individuals. Further research is warranted to determine the relationship between periodontal herpesvirus counts and the risk of viral transmission to close acquaintances.     

Actinobacillus actinomycetemcomitans in human periodontal disease. Prevalence in patient groups and distribution of biotypes and serotypes within families

Actinobacillus actinomycetemcomitans is a Gram-negative oral bacterium which has been implicated in the etiology of localized juvenile periodontitis. In this study, 403 subjects from four study groups were examined for A actinomycetemcomitans in subgingival dental plaque. Samples pooled from at least six periodontal sites were included from each subject. A actinomycetemcomitans was detected in 28 of 29 localized juvenile periodontitis patients but in only 15% of the other subjects including 28 of 134 adult periodontitis patients, 24 of 142 periodontally healthy subjects and 5 of 98 insulin dependent juvenile diabetics with varying degrees of gingivitis. A actinomycetemcomitans isolates from members of five families with localized juvenile periodontitis patients were biotyped on the basis of variable fermentation of dextrin, maltose, mannitol and xylose and serotyped by indirect immunofluorescence using serotype specific rabbit antisera. Individuals within a family all harbored A actinomycetemcomitans of the same biotype and serotype. However, even in families with individuals heavily infected with A actinomycetemcomitans, some family members did not appear to be infected with the organism. The apparent poor transmissibility of A actinomycetemcomitans between individuals may, in part, explain the overall low prevalence of localized juvenile periodontitis and the familial pattern of the disease. The high prevalence of A actinomycetemcomitans in the subgingival plaque of localized juvenile periodontitis patients, compared to the much lower prevalence in other patient groups, supports the hypothesis that A actinomycetemcomitans is an etiologic agent in this periodontal disease.(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of certain subgingival microbial species in selected periodontal conditions

Nine commonly encountered subgingival species were enumerated in subgingival plaque samples from periodontally healthy, gingivitis, and adult and juvenile periodontitis subjects employing two elective and three selective media. Samples were also obtained for darkfield microscopy. Results indicated that Eikenella corrodens and Fusobacterium nucleatum were usually elevated in proportions in sites with gingivitis or destructive periodontal disease. Capnocytophaga gingivalis was associated with gingivitis whereas Capnocytophaga ochracea was found in higher proportions in subgingival plaques of subjects with juvenile periodontitis. Previous association of Actinobacillus actinomycetemcomitans with juvenile periodontitis was confirmed. Spirochetes and other motile organisms were found more frequently and in higher proportions in the three disease states and were very strongly correlated with pocket depth. Motile organisms were also positively correlated with levels of plaque and redness in contrast to cocci which showed a strong negative correlation with all clinical parameters recorded.