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

Abstract

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 o 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.

Salivary Biomarkers: Toward Future Clinical and Diagnostic Utilities

“…Traditional diagnosis and monitoring of HBV and HCV infections consist mainly of blood-based and serological tests evaluating viral load as well as viral antibodies and antigens. Interestingly, reports have indicated that HBV and HCV DNAs, antibodies, and viral antigens not only exist in the saliva of infected subjects but also correlate well with blood samples (75,76,77). These findings suggest a potential role for saliva as a noninvasive mode of HBV and HCV diagnosis and disease state monitoring.…”

Oral Biofluid Biomarker Research: Current Status and Emerging Frontiers

“…Consequently, such epidemiologic investigations can allow researchers to assess risk of disease based on HOMIM-based targeted characterization by comparing to already databased phyla or genera. Infectious diseases, local diseases, and systemic diseases that have been identified are HIV-1, HIV-2, hepatitis A, hepatitis B, hepatitis C, malaria, dengue fever, tuberculosis, Ebola virus disease, herpes simplex, disease caused by and correlated to Epstein-Barr virus, herpesviruses infection, cytomegalovirus-caused illnesses, oral cancer, Crohn’s disease, pancreatic cancer and chronic pancreatitis, periodontal disease, dental caries, and obesity [53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72]. …”

On the origin and diagnostic use of salivary RNA

“…Salivary transcripts can also be used to detect viruses in the oral cavity. The most studied virus for diagnosis associated with salivary RNA is hepatitis C (Wang et al , ; Young et al , ; Chen et al , ; Kage et al , ; Rey et al , ; Goncalves et al , ). Other studied viruses include hepatitis A (Mackiewicz et al , ; Amado et al , ) and immunodeficiency virus‐1 (Kakizawa et al , ; Shugars et al , , ).…”