Human cytomegalovirus and Epstein-Barr virus type 1 in periodontal abscesses

OBJECTIVES: Recent studies have linked herpesviruses to severe types of periodontal disease, but no information exists on their relationship to periodontal abscesses. The present study determined the presence of human cytomegalovirus (HCMV) and Epstein-Barr virus type 1 (EBV-1) in periodontal abscesses and the effect of treatment on the subgingival occurrence of these viruses. MATERIAL AND METHODS: Eighteen adults with periodontal abscesses participated in the study. Subgingival samples were collected from each patient with sterile curettes from an abscess-affected site and a healthy control site. HCMV and EBV-1 were identified by polymerase chain reaction at the time of the abscess and at 4 months after surgical and systemic doxycycline therapy. RESULTS: HCMV was detected in 66.7% of periodontal abscess sites and in 5.6% of healthy sites (P=0.002). EBV-1 occurred in 72.2% of abscess sites but not in any healthy site (P<0.001). HCMV and EBV-1 co-infection was identified in 55.6% of the abscess sites. Posttreatment, HCMV and EBV-1 were not found in any study site. CONCLUSIONS: HCMV and EBV-1 genomes are commonly found in periodontal abscesses. These data favor a model in which a herpesvirus infection of the periodontium impairs the host defense and serves as a platform for the entrance of bacterial pathogens into gingival tissue with subsequent risk of abscess development.     

Human cytomegalovirus and Epstein‐Barr virus type 1 in periodontal abscesses

Objectives: Recent studies have linked herpesviruses to severe types of periodontal disease, but no information exists on their relationship to periodontal abscesses. The present study determined the presence of human cytomegalovirus (HCMV) and Epstein‐Barr virus type 1 (EBV‐1) in periodontal abscesses and the effect of treatment on the subgingival occurrence of these viruses. Material and methods: Eighteen adults with periodontal abscesses participated in the study. Subgingival samples were collected from each patient with sterile curettes from an abscess‐affected site and a healthy control site. HCMV and EBV‐1 were identifed by polymerase chain reaction at the time of the abscess and at 4 months after surgical and systemic doxycycline therapy. Results: HCMV was detected in 66.7% of periodontal abscess sites and in 5.6% of healthy sites (P = 0.002). EBV‐1 occurred in 72.2% of abscess sites but not in any healthy site (P < 0.001). HCMV and EBV‐1 co‐infection was identified in 55.6% of the abscess sites. Posttreatment, HCMV and EBV‐1 were not found in any study site. Conclusions: HCMV and EBV‐1 genomes are commonly found in periodontal abscesses. These data favor a model in which a herpesvirus infection of the periodontium impairs the host defense and serves as a platform for the entrance of bacterial pathogens into gingival tissue with subsequent risk of abscess development.     

Update on human cytomegalovirus in destructive periodontal disease

AIM: Human cytomegalovirus (HCMV), a herpesvirus, is discussed in this review as it relates to destructive periodontal disease in humans. RESULTS: HCMV genomic sequences, detected by polymerase chain reaction identification, occur with elevated frequency in severe adult periodontitis, localized and generalized aggressive (juvenile) periodontitis, Papillon-Lefèvre syndrome periodontitis, acute necrotizing ulcerative gingivitis, and periodontal abscesses. DISCUSSION: Herpesviruses establish lifelong persistent infections. HCMV infection involves an asymptomatic latent phase interrupted by periods of recrudescence where viral replication and possibly clinical disease become manifest. HCMV reactivation is triggered by a number of immunosuppressive factors, some of which have been shown also to be risk factors/indicators of periodontitis. HCMV periodontal infection may cause release of tissue-destructive cytokines, overgrowth of pathogenic periodontal bacteria, and initiation of cytotoxic or immunopathologic events. CONCLUSIONS: A growing body of data supports the concept that HCMV contributes to severe types of periodontal disease. HCMV infection of the periodontium may alter the immune control of resident microorganisms and be important in a multistage pathogenesis of periodontitis involving viral activation, periodontopathic bacteria, and host immune responses. Understanding the significance of HCMV and other herpesviruses in the development of periodontal disease may have important therapeutic implications. Vaccines against HCMV, which are in various stages of development, need to be evaluated for their ability to decrease the incidence of destructive periodontal disease.     

Correlation of cytomegalovirus and human herpesvirus 7 with CD3+ and CD3+ CD4+ cells in chronic periodontitis patients

Thomasini RL, Bonon SH, Durante P, Costa SCB. Correlation of cytomegalovirus and human herpesvirus 7 with CD3 ⁺ and CD3 ⁺ CD4 ⁺ cells in chronic periodontitis patients. J Periodont Res 2012; 47: 114–120. © 2011 John Wiley & Sons A/S Background and Objective: Human chronic periodontitis is an inflammatory process characterized by dense accumulation of immune cells in the periodontal tissue. The periodontitis can lead to loss of teeth in the patient and the pathogenesis of this disease is not completely known. This study tested the hypothesis that chronic periodontitis‐affected sites can harbor betaherpesviruses and that viruses are linked to a profile of the inflammatory infiltrate. Material and Methods: Biopsies of periodontal tissue were taken from periodontitis‐affected patients and from healthy subjects. Immunohistochemistry was performed to count CD19⁺ B cells, CD3⁺ total T cells, T‐CD4⁺ and T‐CD8⁺ cell subsets, and PCR was performed to detect cytomegalovirus and human herpesvirus 6 and 7 in the samples. One slide of each sample was stained with Giemsa for histopathological examination and to evaluate the quality of the cellular infiltrate. Results: As expected, tissues collected from healthy subjects presented no significant level of inflammatory infiltration and were therefore excluded from immunostaining procedures. Results showed that CD19⁺ B cells were in higher number than CD3⁺ T cells in the periodontitis‐affected tissue, but this was not statistically significant. The T‐CD4⁺ lymphocyte subset was significantly higher than the T‐CD8⁺ lymphocyte subset (p = 0.004) in the samples. Cytomegalovirus and human herpesvirus 7 were found at periodontitis‐affected sites, but not in tissue collected from healthy subjects (p = 0.04 and p = 0.04, respectively). Human herpesvirus 6 was rarely detected. We found a correlation between cytomegalovirus and lower CD19⁺/CD3⁺ ratios (ratio < 0.9, p = 0.003) and between human herpesvirus 7 and lower CD19⁺/CD3⁺ ratios (ratio < 0.9, p = 0.003) and higher CD4⁺/CD8⁺ ratios (ratio > 1.1, p = 0.002). Conclusion: This study shows that cytomegalovirus and human herpesvirus 7 can be present at periodontitis‐affected sites but are uncommon at healthy periodontal sites. Moreover, our data suggest that cytomegalovirus can be related to an inflammatory infiltrate with predominance of CD3⁺ T cells, whereas human herpesvirus 7 can be associated with an infiltrate with predominance of T‐CD4⁺ cells. However, further studies are necessary to support this hypothesis. Herpesviruses could play a role in human chronic periodontitis by modulation of the T cell response.     

Herpesviruses in human periodontal disease

Recent studies have identified various herpesviruses in human periodontal disease. Epstein–Barr virus type 1 (EBV‐1) infects periodontal B‐lymphocytes and human cytomegalovirus (HCMV) infects periodontal monocytes/macrophages and T‐lymphocytes. EBV‐1, HCMV and other herpesviruses are present more frequently in periodontitis lesions and acute necrotizing ulcerative gingivitis‐lesions than in gingivitis or periodontally healthy sites. Reactivation of HCMV in periodontitis lesions tends to be associated with progressing periodontal disease. Herpesvirus‐associated periodontitis lesions harbor elevated levels of periodontopathic bacteria, including Acrinobacillus actinomycetemcomitans , Porphyromonas gingivalis , Bacteriodes forsythus , Prevotella intermedia , Prevotella nigrescens and Treponema denticola . It may be that active periodontal herpesvirus infection impairs periodontal defenses, thereby permitting subgingival overgrowth of periodontopathic bacteria. Alteration between latent and active herpesvirus infection in the periodontium might lead to transient local immunosuppression and explain in part the episodic progressive nature of human periodontitis. Tissue tropism of herpesvirus infections might help explain the localized pattern of tissue destruction in periodontitis. Absence of herpesvirus infection or viral reactivation might explain why some individuals carry periodontopathic bacteria while still maintaining periodontal health. Further studies are warranted to delineate whether the proposed herpesvirus‐periodontopathic bacteria model might account for some of the pathogenic features of human periodontal disease.     

Correlation between different genotypes of human cytomegalovirus and Epstein-Barr virus and peri-implant tissue status

Background: The purpose of this study was to estimate the prevalence of different genotypes of human cytomegalovirus (HCMV) and Epstein‐Barr virus (EBV) in peri‐implantitis and mucositis sites, and to evaluate the correlation between herpesvirus presence and clinical parameters. Methods: A total of 80 dental implants (mean time of loading, 4.16 ± 1.8 years) were evaluated during the course of the study (30 peri‐implantitis, 25 mucositis and 25 healthy peri‐implant sites). The following clinical parameters were assessed: visible plaque index, bleeding on probing, suppuration and probing depth. A polymerase chain reaction (PCR) assay was used to identify the presence of different HCMV and EBV genotypes in peri‐implant tissue plaque samples. Results: HCMV‐2 was detected in 53.3% and EBV‐1 in 46.6% of the 30 peri‐implantitis sites evaluated. By contrast, HCMV‐2 was not detected in healthy periodontal sites and EBV‐1 was detected in one healthy site. A statistically significant correlation was found between the presence of HCMV‐2 and EBV‐1 genotypes and clinical parameters of peri‐implantitis. Conclusions: The results from the present study confirmed the high prevalence of HCMV‐2 and EBV‐1 in the peri‐implant tissue plaque of peri‐implantitis sites and suggests a possible active pathogenic role of the viruses in peri‐implantitis.     

Human cytomegalovirus in peripheral giant cell granuloma

Background:  The peripheral giant cell granuloma is a relatively common non-neoplastic inflammatory lesion of gingiva, but the etiopathogeny remains unknown. This study aimed to evaluate the importance of human cytomegalovirus and Epstein–Barr virus in a peripheral giant cell granuloma of a 47-year-old female.
Methods:  The lesion was studied clinically, histopathologically, immunologically and virologically using established procedures.
Results:  The gingival growth was located at the mesial surface of the maxillary left canine having a vital pulp. The mass was 12 × 21 mm in size and exhibited a smooth surface with no evidence of fluctuation on palpation. An excisional biopsy revealed giant cells in a fibrohistiocytic stroma with areas of haemorrhage. Serum protein levels and lymphocyte subsets were within normal limits, except CD3⁺ and CD4⁺ cells were below normal ranges. Polymorphonuclear leukocytes expressed p150,95 (CD11c/CD18) and CXCR-2 receptors within normal ranges, but the CXCR1 receptor showed decreased density, and CD15 were below normal range. A virological sample of the tooth surface adjacent to the gingival swelling yielded 7.6 × 10³ copy-counts of cytomegalovirus and 4.3 × 10³ copy-counts of Epstein–Barr virus.
Conclusions:  The clinical and histological findings were consistent with the diagnosis of peripheral giant cell granuloma. Cytomegalovirus has the potential to induce multinucleated giant cells, and the possibility that the virus contribute to the development of peripheral giant cell granuloma merits further study.     

Real-time polymerase chain reaction quantification of human cytomegalovirus and Epstein-Barr virus in periodontal pockets and the adjacent gingiva of periodontitis lesions

BACKGROUND: Genomic sequences of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV), two herpesviruses, can frequently be detected in periodontal pockets of progressive periodontitis lesions, but the prevalence and load of the two viruses in gingival tissue are unknown. This study determined levels of HCMV and EBV DNA in the periodontal pocket and in the adjacent gingiva of periodontitis lesions using a real-time polymerase chain reaction (PCR) assay. MATERIAL AND METHODS: A total of 20 systemically healthy periodontitis patients participated in the study. Nine patients below 35 years of age were tentatively diagnosed as having aggressive (early onset) periodontitis, and 11 patients 36-56 years of age as having chronic (adult) periodontitis. Clinical parameters were evaluated using established methods. Using periodontal curettes, specimens were harvested from 6-10 mm periodontal pockets and from the adjacent inflamed periodontal pocket wall. A 5'-nuclease (TaqMan) real-time PCR assay was used to identify and quantify genomic copies of periodontal HCMV and EBV. RESULTS: HCMV DNA was detected in 78% of subgingival and 33% of gingival tissue samples from aggressive periodontitis lesions, but only in 46% of subgingival and 9% of gingival tissue samples from chronic periodontitis lesions. In aggressive periodontitis, HCMV subgingival and gingival tissue counts were positively correlated with periodontal pocket depth and probing attachment loss at sample sites (p6 mm, but none of 14 patients having mean pocket depth at sample teeth相似文献   

Periodontitis lesions are the main source of salivary cytomegalovirus

Background:  Herpesviruses play causal or cooperative roles in childhood infections, tumorigenesis, ulcerogenesis, and periodontitis. Saliva is a common vehicle of herpesvirus horizontal transmission, but the source of salivary herpesviruses remains obscure. To evaluate the significance of periodontal disease in shedding of oral herpesviruses, this study determined the genome-copy counts of human cytomegalovirus (HCMV) and Epstein–Barr virus (EBV) in whole saliva of subjects with periodontitis, gingivitis, or no natural teeth.
Methods:  Whole saliva was collected from 14 periodontitis patients, 15 gingivitis patients and 13 complete denture wearers. The study subjects were systemically healthy and had not received periodontal treatment in the past 3 months. Real-time TaqMan polymerase chain reaction was used to determine the salivary load of HCMV and EBV.
Results:  Salivary HCMV was detected in seven (50%) periodontitis patients, but not in any gingivitis or edentulous subjects ( P  < 0.001). Salivary EBV was detected in 11 (79%) periodontitis patients, in five (33%) gingivitis patients, and in seven (54%) edentulous subjects ( P  = 0.076). Salivary samples showed copy counts of HCMV in the range of 3.3 × 10³–4.2 × 10⁴/ml and of EBV in the range of 3.6 × 10²–1.6 × 10⁹/ml.
Conclusions:  HCMV and EBV are commonly present in the saliva of periodontitis patients. Periodontitis lesions of systemically healthy subjects seem to constitute the main origin of salivary HCMV, but do not comprise the sole source of salivary EBV.     

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.     

Collagenase activity and tissue inhibitor of metalloproteinases-1 (TIMP-1) content in human whole saliva from clinically healthy and periodontally diseased subjects

Total TIMP-1 concentration in whole saliva of periodontally diseased subjects, 137 ± 67 ng/ml (mean ± SD), was clearly lower (p < 0.001) than that of clinically healthy subjects, 273±145, and that of edentulous subjects, 332±121. On the contrary, both active [1.58±0.35 units/ml (mean ± SD)] and total (2.08 ± 0.25) collagenase activities inTIMP-1-free whole saliva of diseased subjects were significantly higher than the activities (0.14 ± 0.14 and 0.50 ± 0.27, respectively) in TIMP-1-free whole saliva of healthy subjects. Most of the total collagenase in whole saliva of healthy subjects consisted of procollagenase, while mainly active collagenase was present in whole saliva from patients with periodontal diseases. Significant reciprocal changes of TIMP-1 and collagenase levels, that is, increase in TIMP-1 concentration and decrease in collagenase activity, were observed after the initial therapy of periodontitis patients.     

Effects of long-term use of antiretroviral therapy on the prevalence of oral Epstein-Barr virus

J Oral Pathol Med (2012) 41 : 249–254 Background: The objectives of this study were to determine (i) the prevalence of oral Epstein–Barr virus (EBV) in HIV‐infected subjects compared to non‐HIV controls and (ii) the effects of long‐term use of antiretroviral therapy (ART) on the prevalence of oral EBV. Methods: A cross‐sectional study was performed in HIV‐infected subjects with and without ART, and non‐HIV individuals. DNA in saliva samples was extracted and used as a template to detect EBV BamH1W and EBNA1 by quantitative polymerase chain reaction. Student t‐test and ANOVA test were performed to determine the prevalence rates among groups. Results: Forty‐nine HIV‐infected subjects: 37 on ART (age range 23–54 year, mean 37 year), 12 not on ART (age range 20–40 year, mean 31 year), and 20 non‐HIV controls (age range 19–53 year, mean 31 year) were enrolled. The numbers of EBV BamH1W in saliva were found to be significantly higher in HIV‐infected subjects than non‐HIV controls (80% vs. 20%, mean = 12 118 vs. 134 copies/10⁵ cells, P < 0.001). HIV‐infected subjects who were on ART had significantly lower numbers of EBV BamH1W than those who were not (mean = 4102 vs. 138 613 copies/10⁵ cells, P = 0.011). The numbers were significantly lower in those who received long‐term ART compared with short‐term (mean = 1401 vs. 11 124 copies/10⁵ cells, P = 0.034). No significant difference was observed between the groups when using EBNA1 primers. Conclusions: Prevalence of oral EBV was significantly higher in HIV‐infected subjects than non‐HIV‐controls. The numbers of the virus were significantly decreased by ART. Long‐term use of ART did not increase oral EBV.     

Detection of cytomegalovirus and Epstein-Barr virus in labial salivary glands in Sjogren's syndrome and non-specific sialadenitis

To investigate the role of herpes viruses in Sjogren's syndrome, minor (labial) salivary gland tissues from Sjogren's syndrome and from non-specific sialadenitis were examined for Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) DNA by the polymerase chain reaction. Almost half of all salivary glands studied contained EBV and/or HCMV. There was, however, no significant difference between the detection of EBV or HCMV in salivary glands from patients with Sjogren's syndrome or non-specific sialadenitis. The findings are consistent with the persistence of EBV and HCMV in minor salivary glands following primary infection, but do not indicate a direct role for either virus in the aetiology of Sjogren's syndrome, and do not exclude reactivation of the viruses in this disease.     

Relative distribution of bacteria at clinically healthy and periodontally diseased sites in humans.

Abstract Samples of subgingival bacteria were collected with a clean curette from two relatively normal and two periodontally diseased sites in each of 12 patients with advanced periodontal disease. The samples were suspended in physiologic saline containing 1 % gelatin and examined within 1 hour by darkfield microscopy. From 100-200 bacteria were classified on a percentage basis into one of the following categories: (1) coccoid cells, (2) straight rods, (3) filaments, (4) fusiforms, (5) curved rods, (6) small spirochetes, (7) medium-sized spirochetes, (8) large spirochetes, and (9) motile rods. For each area sampled the following clinical criteria were also recorded: (1) Gingival Index, (2) Plaque Index, (3) probing depth and (4) gingival fluid flow. For each patient separate mean values were calculated for the normal and the diseased sites. The results indicated that significant differences existed in the microbial flora of clinically normal and diseased sites using a paired t-test comparison (2α= 0.001), with coccoid cells more predominant at normal sites (74.3 % vs. 22.3 %), while at diseased sites motile rods were more frequent (12.7 % vs. 0.3 %), as well as curved rods (1.7 % vs. 0 %), small spirochetes (12.6 % vs. 1.1 %), medium-sized spirochetes (18.5 % vs. 0.5 %) and large spirochetes (6.7 % vs. 0.2 %). The ratio of motile to non-motile cells in the normal was 1:49, whereas at diseased sites the ratio was in the vicinity of 1:1. These results clearly show that a different flora is associated with healthy and periodontally diseased sites in the same patient population and that these differences can be detected by means of a technique which is simple and readily adaptable to a clinical setting. The data obtained in this fashion may be useful in monitoring the effect of various treatment modalities on the periodontal flora and possibly in determining the presence or absence of active disease.     

Human cytomegalovirus and Epstein–Barr virus inhibit oral bacteria‐induced macrophage activation and phagocytosis

Introduction: Periodontal disease is an inflammatory condition caused by periodontal microorganisms. Viruses such as human cytomegalovirus (HCMV) and Epstein–Barr virus (EBV) are associated with certain types of periodontal disease, but their roles in promoting the disease are unclear. Because both viruses infect human macrophages, cells which play key roles in the clearance of pathogenic bacteria, it is likely that the viruses alter the functional capacity of macrophages by inhibiting their defense mechanisms against invading pathogens. Methods: Macrophages preinfected with HCMV or EBV were evaluated following stimulation by selected oral bacteria. Bacteria‐induced macrophage activation was assayed by measuring the levels of tumor necrosis factor‐α (TNF‐α) produced in the media, and phagocytic activity was analysed by a phagocytosis assay with fluorescein isothiocyanate‐labeled bacteria. The virus‐infected macrophages were also subjected to semi‐quantitative polymerase chain reaction to measure the expression of toll‐like receptor 9, which is involved in the activation of phagocytosis‐related pathways. Results: Both HCMV and EBV significantly diminished the TNF‐α production typically induced by oral bacteria, inhibited the phagocytic activity of macrophages, and downregulated the expression of toll‐like receptor 9. Conclusion: Infection by HCMV or EBV inhibits the functional ability of macrophages to respond to bacterial challenge, thereby suggesting their pathogenic role in the development of periodontal disease.     

Detection of human viruses in periodontal pockets using polymerase chain reaction

Even though viruses have been implicated in the etiology of several medical and dental disorders, little or no data are available on the possible involvement of human viruses in the pathogenesis of human periodontal disease. This study investigated the presence of human cytomegalovirus, Epstein-Barr virus, herpes simplex virus, human papillomavirus and human immunodeficiency virus (HIV) in crevicular fluid samples from 30 patients with advanced periodontitis and 26 subjects with gingivitis. Viral identification was performed on direct subgingival samples from 3 diseased sites in each patient using the polymerase chain reaction technique. Seventy-eight percent of advanced periodontitis patients were positive for at least one of the five test viruses. Cytomegalovirus was detected in 60% of the periodontitis patients, Epstein-Barr virus in 30%, herpes simplex virus in 20%, human papillomavirus in 17% and HIV in 7%. Forty percent of the periodontitis patients revealed coinfection by 2 to 5 viruses. Only 31 % of the gingivitis subjects showed a positive viral identification in crevicular fluid, and infected individuals only revealed human cytomegalovirus. This study demonstrated that human viruses may occur in periodontitis lesions with relatively high prevalence. The pathogenetic significance of human viruses in destructive periodontal disease needs to be determined.     

Subgingival Epstein-Barr and cytomegalovirus occurrence in pregnancy gingivitis

Background: Although recent studies focused on the role of human herpesviruses in various types of periodontal disease, there was a lack of information in these reports regarding the role of pregnancy gingivitis. The aim of this study is to determine the correlation between pregnancy and the subgingival virus presence and their relationship with clinical parameters. Methods: Seventy pregnant and 40 non‐pregnant women were examined for gingival and plaque indices, bleeding on probing (BOP), and clinical probing depths (PDs) from the whole dentition. Subgingival plaque samples were obtained from sites showing signs of gingivitis and healthy sites. The polymerase chain reaction methodology was used to detect cytomegalovirus (CMV) and Epstein‐Barr virus (EBV) from plaque samples. Results: Our results show that gingivitis lesions in 27 (38.6%) and 10 (14.3%) pregnant patients were positive for EBV and CMV, respectively. In the non‐pregnant group, EBV and CMV were detected in six (15%) and eight (20%) lesions, respectively. A statistically significant difference (P <0.01) was found between the subgingival occurrence of EBV in the two groups. In gingivitis sites, clinical PDs were affected by gestation (P <0.001) and the occurrence of EBV (P <0.001). In healthy sites, clinical PDs were affected by gestation (P <0.05), and BOP was affected by the occurrence of CMV and EBV (P <0.001). Conclusion: Our data indicate that pregnancy increased the risk of the presence of subgingival EBV in pregnant women by 3.647 times more than in non‐pregnant women.