HCV-RNA In Saliva Research Articles

Evaluation of viral load in saliva from patients with chronic hepatitis C infection

Hepatitis C virus can be detected in blood and other bodily fluids, such as saliva. The aim of this study was to detect and quantify the HCV-RNA in saliva and plasma from patients with chronic hepatitis C infections, as well as check the level of viral load in sex groups (age, ethnicity and virus subtypes). Whole saliva and blood from 70 patients with chronic hepatitis C infections attended at the department of gastroenterology from University Hospital. The HCV-RNA load was performed by qRT-PCR using Sybr Green I master mix. HCV-RNA was detected in 80% (56/70) of patients in saliva and 92.85% (65/70) in plasma. The median of the viral load in the plasma was of 4.87 log10, and in saliva, it was 3.32log10, (p = 0.0005). Female patients and black patients exhibited a negative correlation between the HCV-RNA load in saliva vs. the HCV-RNA load in plasma (r = -0.3172, CI95% -0.6240 to -0.03736, p = 0.0491) and (r = -0.3141; IC95% -0.6069 to -0.05926; p = 0.0209), respectively. HCV-RNA was detected and quantified in saliva samples, and according to the quantification levels, saliva may be a possible transmission source of HCV, particularly in women and people of black ethnicity who develop chronic HCV infections.

Detection of hepatitis C virus RNA in saliva of patients with active infection not associated with periodontal or liver disease severity.

BackgroundHepatitis C virus (HCV) is mainly transmitted by parenteral route, being blood transfusion and intravenous drug use the most frequent risk factors. However, it has been suggested that there are other routes of transmission. There are several studies where HCV RNA has been detected in saliva of patients infected with HCV, and epidemiological studies have proposed the dental treatments as possible risk factors for HCV transmission. The purpose of this study was to detect the presence of HCV RNA in saliva of patients with active infection and associating with periodontal or liver disease.MethodsPatients with quantifiable HCV-RNA in serum were enrolled in the study. Periodontal disease was assessed using the modified gingival index (MGI). Presence of dental plaque was assessed with the use of disclosing tablets. Patients were clinically and laboratory evaluated to identify the stage of liver disease, the HCV RNA was determinate in saliva by nested RT-PCR. To determine associations between different parameters univariate and multivariate analysis were used.ResultsA total of 45 patients were included. Of these patients, 21 (46.6%) had hepatitis, 23 (51.1%) had cirrhosis and one patient (2.4%) presented hepatocellular carcinoma (HCC). Viral loads in serum ranged from 2.31–6.68 log IU/ml with a mean of 5.46 log IU/ml (95% CI 5.23–5.70). HCV RNA was positive in saliva of 29 patients (64.4%) and was not detected in 16 (35.6%). For univariate analysis three independent variables were associated with the detection of HCV-RNA in saliva: gender, viral load and dental plaque and multivariate analysis only one independent variable viral load >5.17 log IU/mL remained significantly associated with the detection of HCV in saliva (p = 0.0002). A statistical difference was observed when viral load was analyzed, log 5.85 IU/mL (95% CI 5.67–6.02) for patients with HCV in saliva vs. log 4.77 IU/mL (95% CI 4.35–5.19) for patients without HCV in saliva (p = 0.0001). The detection of HCV-RNA in saliva was more frequent in patients with relatively high serum viral loads.ConclusionHCV-RNA in saliva was associated with the level of serum viral load but not with periodontal or liver disease severity.

Correlation Between Salivary Anti-HCV Antibodies And HCV RNA in Saliva And Salivary Glands of Patients With Chronic Hepatitis C

This study investigated the correlation between anti-HCV antibodies in saliva and HCV RNA in saliva and salivary glands of patients with chronic hepatitis C. Methods: A total of 131 non-stimulated and 49 stimulated saliva samples were tested for anti-HCV antibodies. Results were compared with HCV RNA detection in saliva and salivary glands. Statistical analysis was done. Results: Antibodies were detected in 26.1% of the saliva samples. In 23.5% of these, HCV RNA was detected. Of the 73.9% of saliva samples with undetectable antibodies, 36.8% were positive for HCV RNA. In 26.6% of the patients with salivary gland samples there were detectible antibodies in the saliva. Of these, 11.8% had HCV RNA in the salivary gland. From the 73.4% cases negative for salivary antibodies, 21.3% were positive for HCV RNA in the salivary gland. Conclusion: There was no correlation between anti-HCV antibodies in saliva and HCV RNA in saliva and salivary glands. Support: FAPEMIG.

Stimulated And Non-Stimulated Salivary Flows Should Be Tested For The Presence Of HCV RNA In Saliva Samples From Patients With Chronic Hepatitis C

Objective: In most of the studies which analyzed the presence of HCV RNA in saliva from patients with chronic hepatitis C only stimulated saliva samples have been used for viral detection. Thus, this study compared the prevalence of HCV RNA in non-stimulated and stimulated salivary flows in patients with chronic hepatitis C. Design: Saliva samples of non-stimulated and stimulated salivary flows from 24 patients were collected, and the HCV RNA was investigated by RT-nested PCR. Data regarding age, gender, risk factors for HCV infection, xerostomia and hyposalivation were also analyzed. Results: The HCV RNA could be detected in 11 (45.8%) non stimulated and in 14 (58.3%) stimulated saliva samples, without statistical significance (p=0.472). However, in 18 (75.0%) patients it was possible to detect the presence of the HCV RNA at least in one of the saliva samples. Six (25.0%) patients complained of xerostomia and nine (37.5%) presented hyposalivation, but in only 3 (12.5%) patients, these conditions could be observed, simultaneously. No significant correlation between the presence of HCV RNA in saliva and age, gender, risk factors for HCV infection, xerostomia and hyposalivation could be identified. Conclusion: Both stimulated and non-stimulated saliva samples must be investigated for the presence of HCV RNA in patients with chronic hepatitis C, to avoid underestimated prevalence of HCV in this group of patients.

HCVclearance from saliva ofHIV–HCV‐coinfected patients on treatment with interferon plus ribavirin

To determine hepatitis C virus (HCV) RNA clearance from blood and saliva of HIV-HCV-coinfected patients undergoing combined therapy with pegylated interferon plus ribavirin (PEG-IFN-RIB). Study group was formed of 60 HIV-infected patients with chronic hepatitis C who were starting treatment with PEG-IFN-RIB. Blood and saliva samples were taken at baseline, at the end of treatment and 24 and 48weeks later. A nested RT-PCR technique was used to detect HCV-RNA in saliva. HCV-RNA was detected in saliva at baseline in 64.7% of patients. Thirty-four patients completed follow-up. The response rate (undetectable HCV-RNA) in blood was 79.4% at the end of treatment; 55.8% at 24weeks after the end of treatment and 50% at 48weeks. HCV was detected in saliva of 13 (38.2%) patients at the end of treatment and in 18 (52.9%) patients at 24 and 48weeks later. Concordance of HCV clearance from blood and saliva reached its maximum value at 48weeks after the end of treatment (odds ratio, 112.51). In HIV-HCV-coinfected patients responders to PEG-IFN-RIB, the salivary glands do not appear to be a sanctuary site for HCV, although viral clearance from saliva may be slower than from blood.

Correlation between salivary anti-HCV antibodies and HCV RNA in saliva and salivary glands of patients with chronic hepatitis C

To investigate the correlation between anti-hepatitis C virus (HCV) antibodies in saliva and detection of HCV RNA in saliva and salivary glands of patients with chronic hepatitis C. A total of 180 samples of saliva (131 non-stimulated and 49 stimulated) from 133 patients with chronic hepatitis C were tested by ELISA for presence of anti-HCV antibodies. Results were compared with the detection of HCV RNA in saliva and salivary glands samples. Pearson's chi-squared and Fisher's exact tests were performed for statistical analysis. Anti-HCV antibodies could be detected in 47/180 (26.1%) saliva samples. In 11/47 (23.5%) of these, HCV RNA was also detected. From the 133/180 (73.9%) saliva samples with undetectable anti-HCV antibodies, 49/133 (36.8%) were positive for HCV RNA at least in one saliva sample. From the 64 patients from whom salivary gland samples were available, 17/64 (26.6%) had detectable anti-HCV antibodies in saliva, from which 2/17 (11.8%) also had HCV RNA in the salivary gland. From the 47/64 (73.4%) cases negative for anti-HCV antibodies in saliva, 10/47 (21.3%) were positive for HCV RNA in salivary gland. Taken together, our results suggest that there is no correlation between the presence of anti-HCV antibodies in saliva and the detection of HCV RNA in saliva and salivary glands in patients with chronic hepatitis C. Nevertheless, as there was a statistically significant difference between detection of anti-HCV antibodies and HCV RNA in stimulated saliva, our study points toward the need for new research on mechanisms of HCV shedding in saliva.

Pilot study for assessment of prevalence of intrafamilial hepatitis C transmission in relation to salivary viral load among infected patients with and without chronic renal failure

HCV-RNA in saliva of HCV patients provides a biological basis for its potential transmission. HCV viremia is particularly high in HCV patients on hemodialysis. This study aimed to evaluate the prevalence of HCV in saliva of HCV patients with and without renal failure, and the possible role of intrafamilial transmission of the virus.Twenty HCV patients were enrolled in this study. They were divided into two groups: 10 HCV-infected patients without renal failure (Group I) and 10 with renal failure (Group II). Detection of HCV-RNA by quantitative RT-PCR in serum and saliva of both groups was done. Thirty-eight family members of both groups were included for the detection of serum HCV antibody. The percentage of the saliva-positive patients for HCV was significantly higher in the renal failure group (70%) than the other group (40%) (p<0.05). There was insignificant statistical difference between the two groups as regards infectivity to their family contacts.Also there was insignificant correlation between the level of viremia and the intra familial transmission with a mean+SD (9,33,250+24,501) in negative relatives and a mean+SD(79,912+26,879) in positive relatives (p>0.05). But a significant correlation was revealed between the level of viremia and saliva positivity, with a mean+SD(12,95,666+1792) in saliva-positive patients and a mean+SD (3,74,465+2150) in saliva-negative patients (p<0.05). There was a highly significant difference between infectivity of HCV saliva-positive patients and saliva-negative patients to their family contacts (p<0.001).Conclusion: Increased percentage of HCV detection in saliva of HCV patients with renal failure on HD may cause spreading of HCV in HD units among RF patients. Also there was increased percentage of interfamilial infectivity among the saliva-positive patients to their relatives and this suggests that saliva might have an infective role.

HEPATITIS B AND C IN DENTISTRY

SUMMARY Viral hepatitis is a major public health problem, occurring endemically in all areas of the world. The prevalence of the disease is influenced by numerous factors which may be able to modulate its onset. The presence of HCV-RNA in saliva provides a biological basis for saliva as a possible source of HCV infection, although it does not necessarily imply transmission. Lock and coworkers (2006) demonstrates a contamination with HCV-RNA of a considerable portion of toothbrushes used by hepatitis C patients. Dentists were in a high risk of contracting this disease due to the procedures and instruments of dental treatment. Patients with liver disorders are of significant interest to the dentist because liver plays a vital role in metabolitic function, including the secretion of bile needed to fat absorption, conversion of sugar to glycogen, excretion of bilirubin, a waste product of hemoglobin metabolism. Impairment of liver function can lead to abnormalities of the metabolism of amino acid, protein, carbohydrates and lipid. Many biochemical function performed by the liver, such as synthesis of coagulation factors and drug metabolism, may be adversely affected. Viral hepatitis is the most common liver disorder and is a major public health problem, occurring endemically in all areas of the world. The prevalence of the disease is influenced by numerous factors which may be able to modulate its onset.

Detection of Hepatitis C Virus-RNA in Saliva from Chronically HCV-Infected Patients

The possibility of the non-parenteral Hepatitis C Virus (HCV) transmission is supported by the demonstration that the actual virus is present in several body fluids, including saliva. From a review of the literature many investigators have found the presence of HCV-RNA in saliva, however, widely contrasting results emerge, with detection rates ranging from 0-100%. To further examine HCV salivary shedding, saliva samples were collected from 46 chronically HCV-infected patients and tested for HCV-RNA and occult blood. Quantification and genotyping of serum HCV-RNA were also carried out for each patient. HCV-RNA was detected in 39.13% of the saliva samples. The viral salivary shedding was significantly related to viraemia levels, serum viral genotype and the presence of salivary occult blood. Our findings indicate that the HCV salivary shedding occurs in about one third of HCV infected patients, but seem to suggest that it is unlikely when the serum viral genotype is 3a. Moreover, blood leakage into the oral cavity is possibly the main source of the salivary HCV-RNA. Although the occurrence of the viral salivary shedding does not necessarily mean that HCV transmission occurs by saliva, our results suggest the need for further investigations into the biological factors possibly involved in HCV mucosal transmission related to both the source and the exposed subjects.

Detection of HCV-RNA in Saliva of HIV-HCV Coinfected Patients

The presence of HCV-RNA in saliva of patients with chronic hepatitis C provides a biological basis for the potential transmission of this virus. HCV viremia is particularly high in HCV-HIV-coinfected patients, which could favor the presence of HCV in their saliva. This study was designed to evaluate the prevalence of HCV in saliva of HCV-HIV-coinfected patients. Stimulated whole saliva was collected from 75 HCV-HIV-coinfected patients and 75 HCV controls. The presence of HCV-RNA in saliva was tested by a highly sensitive noncommercialized nested PCR, and analyzed in relation to demographic, clinical, and analytical variables. HCVRNA was detected in the saliva of 49 (65%) HCV-HIV-coinfected patients and 39 (52%) HCV controls. The presence of HCV in saliva was not related to any of the analyzed variables in HCV-HIV-coinfected patients. In the HCV control group a statistically significant relationship was demonstrated only between the detection of HCV-RNA in saliva and the viral load in peripheral blood (p < 0.001). Our results indicate that there is a trend toward a higher HCV-RNA prevalence in the saliva of HCV-HIV-coinfected patients.

Nitric oxide levels are not changed in saliva of patients infected with hepatitis C virus

The aim of this investigation was to determine nitric oxide metabolite levels in saliva samples from hepatitis C virus-positive patients in an attempt to test the hypothesis if increased levels of nitric oxide metabolites correlates with the presence of HCV-RNA in saliva. Saliva of 39 HCV-positive patients and 13 HCV-negative patients, without clinical or laboratorial evidence of liver disease were tested for nitric oxide metabolites. HCV-RNA was detected in serum and saliva by a RT-PCR method and nitric oxide level was determined by evaluation of its stable degradation products, nitrate and nitrite. No differences were found between the concentration of nitrite in saliva from HCV patients and controls, in despite of the presence or not of HCV RNA in saliva. Patients with HCV and cirrhosis had higher concentrations of nitrite but not significantly different from the control group or the groups of anti-HCV patients without cirrhosis. Increased levels of nitrite were not detected in anti-HCV positive patients, an indirect indication that chronic sialoadenitis are infrequent in these patients or occurs with low intensity not sufficient to increase nitric oxide metabolite levels in saliva.

HCV clearance patterns in saliva and serum of patients with chronic HCV infection under interferon plus ribavirin therapy

Hepatitis C virus (HCV)-RNA is often present in saliva of HCV-infected patients, with plasma viral load being the only known predictable factor. Interferon plus ribavirin therapy yields a sustained reduction in HCV viremia. This study aimed to assess the presence of HCV in saliva and serum specimens from patients undergoing this combination therapy (CT). Paired serum and saliva specimens were collected from 44 chronic HCV-infected patients at basal time, 4 and 12 weeks after CT onset, at the end of treatment and 6 months latter. Serum HCV-RNA levels were determined by the polymerase chain reaction (PCR) Amplicor system. Presence of HCV-RNA in saliva was tested by a highly sensitive non-commercialized nested-PCR. The HCV-RNA was detected in 26 saliva specimens at basal time (59.1%). In 34.1% of cases, a concordance viral clearance pattern in serum and saliva was observed in both responders (pattern 1a) and non-responders (pattern 1b). In pattern 2 (13.6% of cases), HCV was detected longer during CT in serum than in saliva (pattern 2a) or in saliva than in serum (pattern 2b). In 11.3% of patients, viral clearance was corroborated either in their serum (pattern 3a) or in their saliva (pattern 3b), but not in both fluids. Of the eight primary responders with 1a clearance pattern, seven were sustained responders. None of the patients with 2a clearance pattern was a sustained responder. Of the two primary responders showing the 3b salivary pattern, one had already relapsed in the first 6 months of follow up. The present results suggest that the monitoring of salivary levels of HCV would be a helpful means of determining sustained antiviral effects of interferon and ribavirin in the treatment of HCV disease.

Detection of hepatitis C virus RNA in saliva samples from patients with seric anti-HCV antibodies

We examined the frequency of HCV-RNA in saliva samples from anti-HCV positive patients. Both plasma and saliva samples from 39 HCV patients (13 with normal liver enzymes, 19 with abnormal liver enzymes and 13 with cirrhosis) were investigated. Stimulated saliva and fresh plasma were centrifuged (900 x g,10 min) and stored at -70 degrees C, after the addition of guanidine isothiocyanate RNA extraction buffer. HCV-RNA was detected by RT-nested-PCR (amplification of HCV-cDNA for two rounds, using HCV primers 939/209 and 940/211). HCV genotyping was carried out by RFLP (using Mva I and Hinf 1 or Hae III and Rsa I restriction enzymes). Thirty-two out of 39 (82%; 95% CI=70-94%) anti-HCV-positive patients had HCV-RNA in plasma samples. Eight out of 39 (20.5%; 95% CI=6.6-34.4%) had HCV-RNA in the saliva. The HCV genotype in saliva samples from these patients matched the genotype found for plasma HCV-RNA. No significant correlation between the presence of HCV and either age, gender, HCV genotype or any risk factor for HCV infection was found. The observed prevalence (20.5% of anti HCV positive patients or 25% of the patients with HCV-RNA in plasma) was lower than that previously reported from other countries. The low frequency of HCV-RNA in saliva samples observed in our study may be due to the use of cell-free saliva. Other authors reporting higher frequencies of HCV-RNA in saliva used whole saliva, without centrifugation.

Detection of hepatitis C virus RNA in saliva is not related to oral health status or viral load

Hepatitis C is a worldwide public health problem and its transmission is clearly associated with the parenteral route, however, the virus has also been isolated from other body fluids. Hepatitis C virus (HCV) RNA has been detected in saliva, yet the relationship between HCV and oral pathology is not clearly understood. Therefore, an investigation on HCV-RNA in saliva and its correlation with oral pathology was undertaken. Saliva and blood samples were collected from 50 anti-HCV positive patients and from 25 patients with non-HCV chronic liver disease. HCV-RNA was detected in all of the saliva samples from the HCV positive group. None of the saliva or serum samples from the non-HCV group were positive for HCV-RNA. The patients were examined for dental and oral health (dentate, partially dentate, edentulous, evidence of gum disease, or mucosal lesions); however, no correlation was found between HCV-RNA in saliva, oral health, and viral load. These results suggest that HCV-RNA presence in saliva is independent of the viral load and the oral pathology of HCV positive individuals.

Detection of HCV-RNA in saliva of patients with chronic hepatitis C virus infection

Detection of HCV-RNA in saliva of patients with chronic hepatitis C virus infection

High prevalence of HCV-RNA in the saliva cell fraction of patients with chronic hepatitis C but no evidence of HCV transmission among sexual partners.

The aims of this study were to evaluate the prevalence of HCV-RNA in different fractions of saliva taken from patients with chronic hepatitis C, to establish whether virologic parameters or disease severity exert any influence on the detectability of HCV-RNA in saliva, and to evaluate the prevalence of HCV infection in partners of HCV-infected subjects with respect to the presence of HCV-RNA in saliva. Sera samples and different fractions of saliva (whole saliva, surnatant, and cell fraction) from 48 subjects (45 with chronic hepatitis C and three healthy anti-HCV+ carriers) were examined for HCV-RNA by RT nested PCR and DEIA hybridization. HCV-RNA-positive sera were also tested for genotype and viral titer (bDNA2 method). Twenty-seven stable sexual partners (25 females and 2 males) were screened for anti-HCV antibodies at least twice over a minimum of 12 months. HCV-RNA was detected in the sera of 39/45 patients and of 22/39 viremic patients. In all of the latter, the presence of HCV-RNA was restricted to the cell fraction. Viral titer was significantly higher in patients with HCV-RNA in saliva than in those without (12.3 x 10(6) versus 4.6 x 10(6) eq/ml, P < 0.01). HCV-RNA positivity was unrelated to genotype, duration of disease, Hepatitis Activity Index scores or transaminase levels. Anti-HCV was positive in one of 13 sexual partners of patients with HCV-RNA in saliva and in 1/14 of those without (P = NS). In conclusion, HCV-RNA is detectable in the cell fraction of saliva in a high proportion of highly viremic patients with chronic hepatitis C, but its presence does not seem to be associated with an increased risk of HCV transmission among sexual partners.

Hepatitis C virus infection of salivary gland epithelial cells: Lack of evidence

Background: Hepatitis C virus genome (HCV-RNA) has been detected in whole salivary gland tissue of chronically infected patients. However, contamination of the tissue by plasma or blood cells was not excluded by the previous reports. Aims: To assess whether HCV infects the salivary gland epithelial cells in patients with chronic HCV liver disease. Methods: Twenty unselected patients with chronic active hepatitis (11 cases) or active cirrhosis (nine cases) were examined. Serum and saliva samples were obtained from all patients, 12 of whom (seven, chronic active hepatitis; five, active cirrhosis) underwent salivary gland biopsy. PCR for HCV-RNA was performed on RNA extracted from serum, saliva and salivary gland epithelial cells collected by isokinetic gradient separation after trypsin digestion of whole salivary gland tissue. Saliva samples were also examined for the presence of secretory IgA anti-HCV by gel chromatography and ELISA testing. Results: HCV-RNA was detected in all sera with titers ranging from 5.42×10 5 genome equivalents/ml to 123.2×10 5 genome equivalents/ml. Thirteen patients were infected with genotype 1b, four patients had genotype 1a, two patients had genotype 2a and one patient was unclassifiable. Low titer HCV-RNA (<2×10 5 genome equivalents/ml) was detected in 3 20 saliva samples (15%) from highly viremic patients infected with 1b genotype. RNA extracted from salivary gland epithelial cells consistently tested negative for HCV-RNA. In addition, all saliva specimens tested negative for secretory-IgA (S-IgA) anti-HCV, even after a 10-fold concentration of the samples. Conclusions: There was no evidence that HCV infects the salivary gland epithelial cells in our viremic patients with HCV chronic liver disease. Low level HCV-RNA in saliva is most probably due to virus spillover from blood.

Detection of HCV-RNA in saliva of patients with chronic hepatitis C.

Previous studies have provided conflicting results on the presence of hepatitis C virus-RNA in saliva. In this study, 23 (62%) of 37 patients tested positive for hepatitis C virus-RNA in saliva, using polymerase chain reaction analysis. A slightly greater proportion had a sporadic rather than a parenteral origin of chronic hepatitis C. These results provide a biological basis for saliva as a possible source of hepatitis C virus (HCV) infection, but do not necessarily imply transmission by this route.