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Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the IIPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 +/- 324,244 copy/mL) than in the HPV-patients (10,246 +/- 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with IIPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

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Evidencias sugieren que los virus pueden participar en la activación de la enfermedad periodontal, permitiendo el sobrecrecimiento de bacterias periodontopatógenas. El objetivo del estudio fué la detección molecular de VPH en fluido gingival (FG) de pacientes VIH+ con enfermedad periodontal. Se evaluaron muestras de FG de 20 pacientes VIH+ con enfermedad periodontal que asistieron al Centro de Atención de Pacientes con Enfermedades Infecciosas (CAPEI) de la Facultad de Odontología de la Universidad Central de Venezuela, 13 bajo terapia antirretroviral (HAART) y 7 VIH+ sin HAART. Se incluyeron 7 pacientes seronegativos con periodontitis crónica y como grupo control 7 pacientes seronegativos periodontalmente sanos. Se extrajo el ADN, se amplificó la región L1 de VPH con primers MY09 y MY11. Las muestras VPH+ fueron genotipificadas para los tipos 6, 11, 16, 18, 31 y 45. VPH fue detectado en 46% de los pacientes VIH+ bajo terapia. El contaje CD4+ en la población VPH+ no presentó diferencias con el grupo VPH-, y la carga viral mostró valores promedio significativamente mayores (200.470± 324.244 copias/mL) con respecto a los pacientes VPH- (10.246±23.805 copias/mL). Las muestras VPH+ presentaron los genotipos 6 y 11, de los cuales 66,6% estaban coinfectados con ambos tipos. Las condiciones periodontales no presentaron diferencias entre los individuos con doble infección viral por VPH y VIH, y los que solo portaban VIH. VPH fue detectado solamente en fluido gingival de pacientes VIH+ con HAART, indicando que esta terapia puede influir en el estado inmunológico independientemente de las condiciones periodontales.

Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the HPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 ± 324,244 copy/mL) than in the HPV- patients (10,246 ± 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with HPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

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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 x 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.

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INTRODUCTION: Chronic graft-vs-host disease (cGVHD) is a major cause of morbidity in long-term survivors of allogeneic hematopoietic progenitor cell transplantation. Herpesviruses are involved in the occurrence and progression of various oral diseases. AIM: The aim of this study was to investigate the role of human herpesvirus 6 (HHV6) in patients with oral manifestations of cGVHD. MATERIALS AND METHODS: Peripheral blood and oral fluids (whole saliva, gingival crevicular fluid and parotid gland saliva) from 19 cGVHD patients, and 28 blood donors were examined for HHV6. Oral tissue samples were collected from 12 cGVHD patients and 12 healthy individuals. Nested polymerase chain reaction was employed to identify the HHV6. RESULTS AND CONCLUSION: The virus was detected in whole saliva in 13 cGVHD patients (68%) and in 19 blood donors (67%). HHV6 was not identified in any of the gingival crevicular fluid and parotid gland saliva samples in cGVHD patients. In the control group 14.3% of both, four gingival crevicular fluid and four parotid gland saliva samples were positive. Two oral tissue samples of cGVHD patients were positive for HHV6. These results indicate that patients with oral manifestations of cGVHD and healthy individuals present high and similar incidence of HHV6 in blood and oral fluids. These data do not support the importance of HHV6 in oral lesions of cGVHD.

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BACKGROUND: Human herpesvirus 6 (HHV6) is the etiologic agent of exanthem subitum. The virus is latent in salivary glands and saliva is the main form of viral transmission. The objective of this study was to assess HHV6 incidence in the fluids from healthy individuals using a standardised technique for collecting and extracting viral DNA from gingival crevicular fluid, whole saliva and parotid gland saliva. DESIGN: Samples of oral fluids and peripheral blood were collected from 28 blood donors and HHV6 was detected using PCR assay. Parotid gland saliva and gingival crevicular fluid were collected by endodontic paper cones in order to not contaminate these fluids with whole saliva. RESULTS: Of the 28 donors, 20 (71.4%) presented positive results in at least one of the three oral fluids researched. Whole saliva was positive in 19 (67.8%) volunteers, while only four (14.2%) samples of gingival crevicular fluid and four of parotid gland saliva proved to be positive. CONCLUSIONS: The results suggest that HHV6 is present in the saliva of a large proportion of the healthy adult population. The use of endodontic paper cones for oral fluid collection and viral extraction was efficient, simple, cheap and painless. In spite of, the small number of cases studied it was possible to demonstrate that neither gingival crevicular fluid nor parotid gland saliva were the principal source of HHV6 in whole saliva.

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