Real-time polymerase chain reaction to determine the prevalence and copy number of epstein-barr virus and cytomegalovirus DNA in subgingival plaque at individual healthy and periodontal disease sites.
ABSTRACT Detection of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) in plaque from patients with periodontal disease provides support for the theory that these viruses play a role in the pathogenesis of periodontitis. This study sought to further define this relationship by determining the prevalence of these viruses at individual disease and healthy sites of patients with periodontal disease and to determine whether the presence and amount of viral DNA correlate with disease severity.
Subgingival plaque from three healthy and three disease sites of 65 patients who had chronic periodontitis were evaluated for the presence and amount of EBV, CMV, and Fusobacterium nucleatum DNA using real-time polymerase chain reaction. Patient serum was evaluated for antibodies against EBV and CMV using enzyme-linked immunosorbent assays.
EBV DNA was detected in 18.5% of subgingival plaque samples (72/390) and in at least one of the six plaque samples in 44.6% (29/65) of the patients. CMV DNA was detected in one plaque sample (0.3%). EBV was significantly more prevalent in disease sites (28.2%; 55/195) than in healthy sites (8.7%; 17/195; P = 0.002). However, neither EBV prevalence nor its amount correlated with increased probing depth >5 mm or attachment loss >2 mm, whereas the amount of F. nucleatum DNA did. Sites positive for EBV had a median copy number of eight. Antibodies against EBV and CMV were detected in 85.7% and 78.6% of persons evaluated, respectively.
EBV was infrequent and CMV was rarely present in individual subgingival sites affected by chronic periodontitis.
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ABSTRACT: Human cytomegalovirus (HCMV) infection is associated with renal transplant failure. Periodontal pockets may be reservoirs for HCMV replication. This study was done to determine active HCMV replication in saliva and gingival crevicular fluid of renal transplant patients affected by periodontitis. HCMV pp67-mRNA amplification was analyzed in oral fluids of 38 transplant recipients at 6 months' posttransplantation. Patients received antiviral therapy until 3 months' posttransplantation. The HCMV-positive cell line VR-977 was the positive control, and oral fluids from healthy volunteers served as the negative control. Periodontitis was diagnosed by clinical examination. Serum HCMV IgG and IgM were analyzed to differentiate recent and latent infection. Prevalence of gingival overgrowth was 68.4%. HCMV gene transcripts were detected in the saliva of 21% and the gingival crevicular fluid of 18% of patients. All patients (100%) with HCMV pp67-mRNA detected in saliva demonstrated clinical manifestations of viral infection, as did 86% of patients with HCMV pp67-mRNA detected in the gingival crevicular fluid. Serum IgM was positive in 7.9% of patients and IgG in 65.8%; however, associations with active mRNA replication were not statistically significant. Renal transplant patients affected by periodontitis are at risk of viral replication within the periodontal tissues despite antiviral therapy. This study suggests that use of HCMV pp67-mRNA detection in saliva and gingival crevicular fluid provides markers of active viral infection, and evidence for a link between HCMV-associated periodontitis and renal transplant complications.Transplantation Proceedings 01/2004; 35(8):2949-52. · 0.95 Impact Factor
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ABSTRACT: Human cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1) are frequently detected in human periodontitis lesions. However, no information is available on the types of gingival cells infected by herpesviruses. The present study determined the presence of herpesviruses in polymorphonuclear neutrophils, monocytes, macrophages and T and B lymphocytes in biopsies of periodontitis lesions from 20 adults. A nested polymerase chain reaction method was employed to detect HCMV, EBV-1, EBV-2, human herpes virus-6 and herpes simplex virus (HSV) in periodontal tissue biopsy and in gingival cell fractions separated by immunomagnetic cell sorting. Tissue specimens from 18 (90%) and cell fractions from 14 (70%) patients demonstrated herpesviruses. Periodontitis-derived monocytes and macrophages revealed HCMV in cell fractions from 11 (55%) patients and HSV in cells from 1 (5%) patient. T lymphocytes harbored HCMV in cell fractions from 4 (20%) patients and HSV in cell fractions from 4 (20%) patients. B lymphocytes showed EBV-1 in cell fractions from 9 (45%) patients. Periodontal polymorphonuclear neutrophils demonstrated no herpesviruses. This study suggests that HCMV mainly infects periodontal monocytes, macrophages and less frequently T lymphocytes and that EBV-1 infects periodontal B lymphocytes. The possible etio-pathologic significance of periodontal herpesvirus infection is discussed.Oral Microbiology and Immunology 09/1999; 14(4):206-12. · 2.81 Impact Factor
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ABSTRACT: Herpesviruses are implicated in the pathogenesis of human periodontitis. However, the quantity of herpesviruses in periodontal sites remains unknown. The aim of this study was to compare levels of subgingival human cytomegalovirus (HCMV) in aggressive periodontitis patients and in periodontally healthy subjects. A total of 16 consecutive subjects with aggressive periodontitis and 15 healthy control subjects were included in the study. Subgingival specimens were collected by a periodontal curette. TaqMan real-time polymerase chain reaction (PCR) assay was used to quantify HCMV. HCMV was detected in 68.8% of aggressive periodontitis lesions but not in any of the periodontally healthy study sites. HCMV viral load in positive subgingival specimens ranged from 5 x 10(2) to 7.4 x 10(3) copies/ml. The TaqMan real-time PCR technology seems to provide a rapid and sensitive method for quantifying HCMV in periodontal pockets. The present findings confirm the frequent presence of HCMV in aggressive periodontitis lesions. Determining HCMV levels in different types of periodontitis may help elucidate the periodontopathic role of the virus and advance our understanding of the disease pathogenesis.Journal of Periodontal Research 05/2004; 39(2):81-6. · 1.99 Impact Factor