HCMV gB genotype and its association with cytokine levels in hematopoietic stem cell transplantation

Oral Diseases (2011) 17 , 530–537 Background: Glycoprotein B (gB) has been implicated in determining the pathogenicity and clinical outcomes of human cytomegalovirus (HCMV) disease. Objective: The purpose of this study was to assess the prevalence of gB genotypes in allogeneic hematopoietic stem cell transplantation (allo‐HSCT) and the relationship between it and cytokine levels in saliva and blood samples. The impact of these parameters on patients’ survival was also investigated. Methods: Samples were obtained from 63 patients receiving an allo‐HSCT. HCMV gB genotyping was carried out by multiplex nested PCR. The cytokine levels were assessed using ELISA assay. Results: A single or mixed genotype infection was detected in the saliva and blood of 36/63 and 52/63 subjects, respectively. Patients with gB2 in their saliva showed lower IL‐10 levels in comparison with patients without gB2. Reduced blood levels of IFN‐γ and IL‐1β were also found in recipients with the HCMV gB4 genotype compared with patients without it. Decreased IL‐1β and increased IL‐10 blood levels were associated with lower survival. However, HCMV gB genotypes have no impact on patient outcome. Conclusion: Decreased IL‐1β and increased IL‐10 levels in the blood are associated with lower survival. HCMV genotypes are associated with different cytokine levels in saliva and blood.     

Comparison of nested polymerase chain reaction (PCR), real‐time PCR and viral culture for the detection of cytomegalovirus in subgingival samples

Introduction: The purpose of this study was to compare nested polymerase chain reaction (PCR), real‐time PCR, and shell vial for the detection of human cytomegalovirus (HCMV) in subgingival samples in periodontitis patients. Methods: A group of 44 patients and 24 individuals without periodontitis were included in the study. A full periodontal examination was conducted in each subject. Gingival crevicular fluid (GCF) was collected by pocket lavage and used for viral culture (shell vial). Additional subgingival samples were obtained with paper points and used for molecular analysis. Nested PCR and real‐time PCR were used to detect and quantify HCMV. Student’s t‐test and chi‐squared test were used to compare groups. The sensitivity and specificity for the tests were calculated on 2 × 2 tables considering the nested PCR as the gold standard. Results: The detection of HCMV was greater using nested PCR than with either real‐time PCR or shell vial (P < 0.0001). However, the frequency detection of both molecular techniques was higher than in viral culture (P < 0.0001). Only one case of chronic periodontitis was positive by viral culture. Agreement between nested PCR and real‐time PCR was observed 47.7% and 4.1% of the time in the periodontitis and control groups, respectively. The sensitivity of real‐time PCR was 60%, compared with 2.8% for the shell vial technique. Conclusions: In conclusion, this study confirmed that active HCMV infection occurs in human periodontitis; however, its frequency seems to be low. In contrast, latent periodontal HCMV infection seems to be a more frequent event.     

Relationship between herpesviruses and periodontopathogens in patients with HIV and periodontitis

Background: The purpose of the present study is to verify a possible association between herpesviruses and periodontal pathogens in individuals with human immunodeficiency virus (HIV) and periodontitis. Methods: Twenty‐seven patients with HIV and chronic periodontitis and 23 patients with HIV and gingivitis were included in the study. Probing depth, clinical attachment loss, gingival index, and plaque index were recorded. Blood, saliva, and subgingival plaque were processed for viral and bacterial identification. Bacteria were identified by 16S rRNA‐based polymerase chain reaction and viruses by the nested polymerase chain reaction. Results: For the chronic periodontitis group, Epstein‐Barr (EBV)‐1 (70.4%) and Tannerella forsythia (Tf) (51.8%) presented higher detection in subgingival plaque and saliva (81.5% and 40.7%, respectively) than in blood (22% and 0%, respectively) (P <0.005 and P <0.0001, respectively). Porphyromonas gingivalis (Pg) was more frequent in subgingival plaque (77.7%; P <0.0001). In the gingivitis group, Pg and human cytomegalovirus (HCMV) presented higher frequency in subgingival plaque (95.6% and 91.3%, respectively; P <0.0001 and P = 0.004). Tf and EBV‐1 were detected more frequently in subgingival plaque (47.8% and 78.3%, respectively) and saliva (52.2% and 52.2%, respectively; P = 0.004 and P <0.005) than in blood. EBV‐1, EBV‐1–HCMV, and presence of different viruses presented an association with periodontitis in saliva. Conclusions: No association was detected for herpesviruses and periodontal pathogens in patients who are HIV‐positive with periodontitis. EBV‐1 and coinfection (EBV‐1–HCMV) were associated with patients who are HIV‐positive with periodontitis.     

Quantitative Bestimmung humaner CMV-DNA im Speichel von Patienten mit Knochenmark- und Stammzelltransplantation mithilfe der TaqMan®-PCR

Background

Human cytomegalovirus (HCMV) infection is associated with severe and life-threatening diseases in immunocompromised patients, especially after bone marrow (BM) and stem cell (SC) transplantation. Prior to transplantation the potential risk of HCMV disease is therefore determined by HCMV-antibody blood testing of transplant donor (D) and recipient (R). Virus carriers are positive for anti-CMV-IgG. Virus patterns are distinguished as follows: group 1 (D+/R+), group 2 (D?/R+), group 3 (D+/R?), and group 4 (D?/R?).

Aim

The aim of this study was qualitative and quantitative determination of the HCMV DNA load in saliva of BM and SC transplantation patients.

Patients and method

Unstimulated saliva was collected from 20 patients prior to BM and SC transplantation, during the time of conditioning, and after transplantation. DNA was isolated and analyzed for evidence of HCMV DNA with TaqMan PCR.

Results

HCMV DNA was isolated in seven cases. In all group 1 patients (D+/R+) HCMV DNA could be demonstrated. Only three of seven group 2 patients (D?/R+) were positive for HCMV DNA. The only group 3 patient (D+/R?) and all eight group 4 patients (D?/R?) were negative.

Conclusion

TaqMan PCR is a reliable method for HCMV DNA quantification. In three patients (anti-HCMV-IgG positive) who received an anti-CMV-IgG negative transplant HCMV DNA was isolated. In contrast, no HCMV-DNA was evident in HCMV-negative patients who received an HCMV-negative transplant. Accordingly, the risk of HCMV reactivation is more probable than the risk of reinfection.      

Saliva and blood interferon gamma levels and IFNG genotypes in acute graft-versus-host disease

Oral Diseases (2012) 18 , 816–822 Objective: Graft‐versus‐host disease is a major complication after allogenic hematopoietic stem cell transplantation. Interferon gamma is an important pro‐inflammatory cytokine involved in this disease. Cytokine gene polymorphisms are associated with functional differences in cytokine expression and can alter the clinical course of graft‐versus‐host disease. This study aimed to investigate the association between IFN‐γ levels in saliva, blood, and IFNG polymorphisms, as well as the occurrence of acute graft‐versus‐host disease in allogenic HSCT. Subjects and Methods: Fifty‐eight consecutive allogenic hematopoietic stem cell transplantation recipients and their donors were prospectively studied. IFN‐g levels in saliva and blood were assessed by ELISA. Samples were collected weekly from 7 days before transplantation (day ?7) to 100 days after allogenic HSCT (day +100) or until death. Saliva and/or blood samples were obtained from the recipients and donors to determine IFNG gene polymorphisms. Results: Increased saliva and blood IFN‐g levels were observed in patients that had developed aGVHD. In the saliva, the peak levels of IFN‐g could be found one week before aGVHD diagnosis, while in the blood, peak levels of IFN‐g could be only observed upon diagnosis. A significant association could be identified between the recipients’IFNG genotypes and the IFN‐g levels in their blood, at +14 days after HSCT. No association could be observed between IFNG gene polymorphisms and the aGVHD. Conclusion: The present study shows that the genetic background of recipients can influence the production of IFN‐g. Moreover, as IFN‐g levels in the saliva and blood were found to be associated with aGVHD development, this cytokine may be a useful predictor of acute graft‐versus‐host disease.     

Evaluation of Biochemical Parameters and Local and Systemic Levels of Osteoactive and B‐Cell Stimulatory Factors in Gestational Diabetes in the Presence or Absence of Gingivitis

Background: Gestational diabetes mellitus (GDM) is defined as varying glucose intolerance, with first onset or recognition in pregnancy. This study evaluates clinical and biochemical parameters in a possible association between GDM and gingivitis. Methods: A total of 167 pregnant females was included in the study. There were 101 females with GDM and 66 females without GDM. Subgroups were created according to the presence or absence of gingival inflammation. Plaque index, bleeding on probing, and probing depth were recorded at four sites per tooth. Serum, saliva, and gingival crevicular fluid (GCF) levels of interleukin (IL)‐6, IL‐8, soluble receptor activator of nuclear factor‐kappa B ligand (sRANKL), osteoprotegerin (OPG), B‐cell activating factor (BAFF), and a proliferation‐inducing ligand (APRIL) were determined by enzyme‐linked immunosorbent assay. Data were analyzed by Kruskal‐Wallis and Mann‐Whitney U tests and Spearman correlation analysis. Results: Age and anthropometric indices were higher in the GDM than non‐GDM group (P <0.0001). Clinical periodontal recordings, serum BAFF, IL‐8, and saliva sRANKL levels were higher in the GDM group (P <0.05). Saliva IL‐6 level was higher in the GDM with gingivitis group than non‐GDM with gingivitis group (P = 0.044). Serum and GCF BAFF (P <0.0001), serum, saliva, and GCF APRIL (P <0.0001; P <0.0001; P = 0.032, respectively), GCF OPG (P = 0.036), and serum and saliva sRANKL (P <0.0001) were higher in the GDM with gingivitis group than GDM without gingivitis group. Conclusions: The inflammatory response seems to be more pronounced in females with GDM. The observed increase in both local and systemic levels of inflammatory cytokines may suggest an interaction between gingivitis and GDM.     

Trefoil factors in saliva and gingival tissues of patients with chronic periodontitis

Background: Trefoil factors (TFFs) are secreted molecules that are involved in cytoprotection against tissue damage and the immune response. TFFs have been detected in saliva and oral tissues, but their clinical significance has never been investigated in patients with chronic periodontitis. The objective of this study is to determine whether TFF expression in saliva and gingival tissues is associated with periodontal pathology. Methods: Saliva and gingival tissue samples were collected from 25 non‐periodontitis individuals and 25 patients with chronic periodontitis (CP). Enzyme‐linked immunosorbent assay and immunohistochemical methods were used to evaluate the expression of TFFs (TFF1, TFF2, and TFF3) in saliva and gingival tissues, respectively. Periodontopathic bacteria were quantified by real‐time polymerase chain reaction. Results: Reduced salivary TFF1 and TFF3 concentrations were observed in patients with CP (P = 0.003 and P <0.001, respectively). Decreased TFF3 expression in gingival tissues of patients with CP was demonstrated (P = 0.041). Levels of salivary TFF3 concentrations were negatively correlated with periodontal pathology and number of Porphyromonas gingivalis and Tannerella forsythia (formerly known as Bacteroides forsythus). Conclusions: Altered expression of TFFs in saliva and gingival tissues was detected in patients with CP. The results suggest that TFF3 may be involved in the pathogenesis of periodontal disease.     

Chitinase expression in parotid glands of non-obese diabetic mice

Oral Diseases (2012) 18 , 506–512 Objective: This investigation was a basal study that used a mouse model of xerostomia to identify protein biomarkers of xerostomia in saliva. We identified genes expressed differently in parotid glands from non‐obese diabetic mice with diabetes and those from control mice; subsequently, we investigated expression of the proteins encoded by these genes in parotid glands and saliva. Materials and Methods: DNA microarray and real‐time PCR analyses were performed to detect differences between NOD/ShiJcl and C57BL/6JJcl (control) female mice in gene expression from parotid glands or parotid acinar cells. Subsequently, protein expression was assessed using immunoblotting and immunohistochemistry. Similarly, enzyme activity in saliva was assessed using zymography. Results: Based on gene expression analyses, Chia expression was higher in diabetic mice than non‐diabetic mice and control mice; similarly, expression of chitinase, the protein encoded by Chia, was higher in diabetic mice. Saliva from NOD/ShiJcl mice had more chitinase than saliva from control mice. Conclusions: Chitinase was highly expressed in parotid acinar cells from diabetic mice compared with non‐diabetic and control mice. Increased chitinase expression and enzyme activity may characterize the autoimmune diabetes in mice; however, further investigation is required to assess its use as a biomarker of xerostomia in humans.     

Salivary shedding of HHV‐8 in people infected or not by human immunodeficiency virus 1

J Oral Pathol Med (2011) 40 : 97–102 Background: Human herpesvirus 8 (HHV‐8), the main agent involved in the etiopathogenesis of Kaposi’s sarcoma (KS) is primarily transmitted through sexual contact. The potential of saliva as a source of HHV‐8 transmission remains unclear. The purpose of this work was to determine the frequency of HHV‐8 detection in saliva of HIV‐infected individuals and their family contacts. Methods: The study group comprised 210 individuals. Group 1: 35 HIV‐infected patients; group 2: 35 non‐HIV individuals; group 3: two siblings for each patient from group 1; group 4: two siblings for each individual from group 2. Each participant had non‐stimulated whole saliva collected and DNA was extracted. HHV‐8‐DNA amplification from ORF‐26 was performed using a nested PCR protocol. Results: HHV‐8 DNA was detected in saliva from 14/35 (40%) HIV‐infected individuals and 4/35 (11.4%) non‐HIV‐infected individuals (OR = 5.16, CI [1.49–17.88], P = 0.006). It was also possible to amplify HHV‐8 DNA in 11/70 (15.7%) relatives of HIV‐infected participants and 4/70 (5.71%) relatives of non‐HIV‐infected individuals(P = 0.041). Among the 14 group 1 patients with HHV‐8 DNA detected in saliva, eight (57.1%) had a household member in whom HHV‐8 DNA was also amplified (OR = 8, CI [1.58–40.29] P = 0.007). Conclusions: HHV‐8 DNA is frequently found in saliva. HIV‐infected individuals showed a higher frequency of detection of HHV‐8 than healthy controls. HHV‐8 DNA was significantly amplified in saliva of household members of HIV/HHV‐8 co‐infected individuals.     

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.     

Evaluation of the long-term storage stability of saliva as a source of human DNA

Objectives

The objectives of this paper are to determine the storage stability of saliva at 37 °C over an 18-month period, and its influence on the DNA yield, purity, PCR protocols and genotyping efficacy.

Materials and methods

Of the 60 participants, blood samples were obtained from 10 and saliva from 50. Samples were subjected to different storage conditions: DNA extracted immediately; DNA extracted following storage at 37 °C for 1, 6, 12 and 18 months. Subsequently, DNA yield, OD_260/280 and OD_260/230 ratios were measured. The isolated DNA was used to amplify exons 0–7 of the RUNX2 gene and subsequently sequenced. Furthermore, 25 SNPs were genotyped.

Results

The mean DNA yield, OD_260/280 and OD_260/230 ratios obtained from blood were 67.4 ng/μl, 1.8?±?0.05 and 1.8?±?0.4 respectively. DNA yield obtained from saliva was significantly higher than blood (p?<?0.0001), ranging from 97.4 to 125.8 ng/μl while the OD_260/280 ratio ranged from 1.8?±?0.13 to 1.9?±?0.1. The success rates for the 25 SNPs ranged from 98 to 100 % for blood and 96–99 % for saliva samples with the genotype frequencies in Hardy–Weinberg equilibrium (>0.01).

Conclusions

Saliva can be stored at 37 °C for 18 months without compromising its quality and ability to endure genetic analyses.

Clinical relevance

Saliva is a viable source of human DNA to facilitate the feasibility of large-scale genetic studies.     

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.     

Herpes viruses in periodontal compromised sites: comparison between HIV-positive and -negative patients

Aim: The objective of this study was to compare the frequency of herpes simplex virus type 1 (HSV‐1), Epstein–Barr virus (EBV) and human cytomegalovirus (HCMV) in subgingival plaque, saliva and peripheral blood of HIV‐positive and‐negative patients with periodontal disease. Material and Methods: Fifty HIV‐positive subjects (23 with gingivitis, 27 with periodontitis) and 50 healthy HIV‐negative patients with chronic periodontitis were included in the study. Parameters of probing depth (PD), clinical attachment level (CAL), gingival index and plaque index were recorded. The samples were processed for viral identification by the nested polymerase chain reaction technique. Results: HCMV was the most prevalent virus in HIV‐positive (82%) and‐negative patients (84%), and the detection in the three samples was similar (p>0.05). HSV‐1 was the least prevalent virus in both groups, being detected in similar frequencies in oral sites and in peripheral blood. EBV‐1 was found more frequently in saliva and subgingival plaque of HIV‐positive patients than in HIV‐negative patients (p0.05). Conclusions: EBV‐1 was more frequently recovered in oral sites of HIV‐positive patients than in HIV‐negative patients.     

Detection and quantification of herpesviruses in Kostmann syndrome periodontitis using real‐time polymerase chain reaction: a case report

Background/aims: Kostmann syndrome, or severe congenital neutropenia, is an autosomal recessive disease of neutrophil production and is associated with severe periodontal pathology. The aim of this study was to determine whether human cytomegalovirus (HCMV) and Epstein‐Barr virus (EBV) contribute to the pathogenesis of Kostmann syndrome periodontitis. Methods: Supragingival plaque and saliva samples were taken from a 6‐year‐old boy and his 3‐year‐old sister suffering from Kostmann syndrome, and from two age‐ and gender‐matched healthy children serving as controls. The samples were taken before and 24 months after periodontal treatment. Real‐time polymerase chain reaction (TaqMan Real‐Time PCR) assay was used to quantify HCMV and EBV DNA. Results: EBV was detected in baseline samples from the Kostmann syndrome patients but not in samples from the healthy control subjects. HCMV was only detected in the saliva of the boy with Kostman syndrome at baseline. Herpesviruses numbers decreased dramatically in the post‐treatment samples. Conclusion: EBV and HCMV were detected in the two subjects with Kostmann syndrome periodontitis. The results of the study indicate that nonsurgical treatment of Kostmann syndrome periodontitis can reduce supragingival and salivary herpes viral loads.     

Immunophenotyping in Saliva as an Alternative Approach for Evaluation of Immunopathogenesis in Chronic Periodontitis

Background: To date, flow cytometric immunophenotyping has not been used to investigate immune patterns in saliva samples from individuals with inflammatory processes in the oral cavity, such as chronic periodontitis (CP). Saliva analysis could be a non‐invasive method for evaluating oral health. The objective of this study is to determine the phenotype of leukocytes and total immunoglobulin A (IgA), IgG, and IgM titers in the saliva of individuals with CP. Methods: Saliva samples were obtained from patients with CP (n = 12) and from a control group (n = 27) without oral diseases. Flow cytometry was performed to determine the frequency of T cells (CD4⁺ and CD8⁺), B cells, and natural killer (NK) cells as well as the total leukocyte population. Immunoglobulin titers were determined by dot enzyme‐linked immunosorbent assay. Results: Cell immunophenotyping revealed that patients with CP had a higher frequency of total leukocytes (47.94% ± 5.1%; P < 0.001), B cells (43.93% ± 6.2%; P = 0.006), NK cells (0.16% ± 0.04%; P = 0.03), and CD4⁺ T cells (38.99% ± 4.4%; P = 0.002) than individuals without oral pathologies (24.75% ± 2.2%, 20.60% ± 2.7%, 0.09% ± 0.03%, and 16.82% ± 3.5%, respectively). No significant differences in salivary total IgA, IgG, and IgM titers were found between the two cohorts studied. Nevertheless, higher total IgG levels were observed in patients with CP, which could indicate a possible correlation between clinical attachment level and salivary IgG (P = 0.07; r² = 0.08). Conclusion: These results show that cell phenotyping by flow cytometry could be an effective tool for determining leukocyte profiles in saliva samples from patients with CP and healthy individuals.     

Comparison of nested polymerase chain reaction (PCR), real-time PCR and viral culture for the detection of cytomegalovirus in subgingival samples

Introduction:  The purpose of this study was to compare nested polymerase chain reaction (PCR), real-time PCR, and shell vial for the detection of human cytomegalovirus (HCMV) in subgingival samples in periodontitis patients.
Methods:  A group of 44 patients and 24 individuals without periodontitis were included in the study. A full periodontal examination was conducted in each subject. Gingival crevicular fluid (GCF) was collected by pocket lavage and used for viral culture (shell vial). Additional subgingival samples were obtained with paper points and used for molecular analysis. Nested PCR and real-time PCR were used to detect and quantify HCMV. Student's t -test and chi-squared test were used to compare groups. The sensitivity and specificity for the tests were calculated on 2 × 2 tables considering the nested PCR as the gold standard.
Results:  The detection of HCMV was greater using nested PCR than with either real-time PCR or shell vial ( P  < 0.0001). However, the frequency detection of both molecular techniques was higher than in viral culture ( P  < 0.0001). Only one case of chronic periodontitis was positive by viral culture. Agreement between nested PCR and real-time PCR was observed 47.7% and 4.1% of the time in the periodontitis and control groups, respectively. The sensitivity of real-time PCR was 60%, compared with 2.8% for the shell vial technique.
Conclusions:  In conclusion, this study confirmed that active HCMV infection occurs in human periodontitis; however, its frequency seems to be low. In contrast, latent periodontal HCMV infection seems to be a more frequent event.     

Comparing culture, real-time PCR and fluorescence resonance energy transfer technology for detection of Porphyromonas gingivalis in patients with or without peri-implant infections

Galassi F, Kaman WE, Anssari Moin D, van der Horst J, Wismeijer D, Crielaard W, Laine ML, Veerman ECI, Bikker FJ, Loos BG. Comparing culture, real‐time PCR and fluorescence resonance energy transfer technology for detection of Porphyromonas gingivalis in patients with or without peri‐implant infections. J Periodont Res 2012; 47: 616–625. © 2012 John Wiley & Sons A/S Background and Objective: The aim of the study was to compare the detection of Porphyromonas gingivalis using a fluorescence resonance energy transfer (FRET) technology with commonly used diagnostic methods in salivary and subgingival plaque samples from subjects with dental implants. P. gingivalis was considered as a marker for a pathogenic microbiota. Material and Methods: Ninety‐seven adult subjects were recruited, including periodontally healthy controls with no dental implants, implant controls with no peri‐implant disease and patients with peri‐implant disease. Saliva and subgingival/submucosal plaque samples were collected from all subjects and were analyzed using culture, real‐time PCR and FRET technology employing P. gingivalis‐specific substrates. Results: It was found that the P. gingivalis‐specific substrates were highly suitable for detecting the presence of P. gingivalis in saliva and in subgingival plaque samples, showing comparable specificity to culture and real‐time PCR. Conclusion: We applied the FRET technology to detect P. gingivalis in implant patients with or without an implant condition and in controls without implants. The technique seems suitable for detection of P. gingivalis in both plaque and saliva samples. However, with all three techniques, P. gingivalis was not very specific for peri‐implantitis cases. Future work includes fine‐tuning the FRET technology and also includes the development of a chair‐side application.     

Detection and quantification of herpesviruses in Kostmann syndrome periodontitis using real-time polymerase chain reaction: a case report

BACKGROUND/AIMS: Kostmann syndrome, or severe congenital neutropenia, is an autosomal recessive disease of neutrophil production and is associated with severe periodontal pathology. The aim of this study was to determine whether human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) contribute to the pathogenesis of Kostmann syndrome periodontitis. METHODS: Supragingival plaque and saliva samples were taken from a 6-year-old boy and his 3-year-old sister suffering from Kostmann syndrome, and from two age- and gender-matched healthy children serving as controls. The samples were taken before and 24 months after periodontal treatment. Real-time polymerase chain reaction (TaqMan Real-Time PCR) assay was used to quantify HCMV and EBV DNA. RESULTS: EBV was detected in baseline samples from the Kostmann syndrome patients but not in samples from the healthy control subjects. HCMV was only detected in the saliva of the boy with Kostman syndrome at baseline. Herpesviruses numbers decreased dramatically in the post-treatment samples. CONCLUSION: EBV and HCMV were detected in the two subjects with Kostmann syndrome periodontitis. The results of the study indicate that nonsurgical treatment of Kostmann syndrome periodontitis can reduce supragingival and salivary herpes viral loads.