Absence of Hepatitis C Virus and Detection of Hepatitis G Virus/GB Virus C RNA Sequences in the Semen of Infected Men

Abstract

and Christian BrechotThe identification of hepatitis C virus (HCV) in semen remains controversial and that of hepatitis G virus (HGV) or GB virus C (GBV-C) has never been investigated. Serum and semen from 90 anti-HCV -positive drug users were tested (27 infected with HIV) for HCV and HGV/GBV-C RNAs by polymerase chain reaction (PCR) assay, hybridization, and sequence analysis. Semen was processed into round cells, seminal plasma, and spermatozoa. Fifty-six patients were HCV-viremic, but HCV-RNA was not identified in their seminal fractions. However, PCR inhibitors were found in the semen of 34 of these men. Twenty-eight patients had HGV/GBV-C RNA in their blood and for 24 of them, ejaculates were available for analysis. HGV/GBV-C RNA was found in the seminal plasma of 6 of 12 samples free from PCR inhibitors. These results agree with the low risk of sexual transfer of HCV and provide preliminary evidence for the presence of HGV/GBV-C in semen. Several investigators have shown that the sex partners ofThe data currently available suggest a low pathogenicity of HGV/GBV-C infection, but recently it has been suggested that subjects infected with hepatitis C virus (HCV) have a low prevalence of anti-HCV antibody, suggesting a low risk of some virus strains might be involved in certain cases of fulminant hepatitis [28, 29]. While this point continues to be debated sexual transmission of this virus [1 -7]. On the contrary, other studies propose that sexual transmission might have a role in [30, 31] it is still important to understand the biology of HGV/ GBV-C infection. Parenteral transmission through contamithe epidemiology of HCV and account for some cases of sporadic HCV-related chronic hepatitis [8 -11]. Studies to identify nated blood has been confirmed, but it is not known whether this virus can be sexually transmitted. HCV RNA in the semen of infected subjects by polymerase chain reaction (PCR) assay have yielded conflicting resultsIn this study, we looked for HCV and HGV/GBV-C RNAs in the serum and semen of 90 anti-HCV -positive IVDUs, of [12 -15], possibly due to difficulties in the extraction of nucleic acids and the presence of PCR inhibitors in seminal plasma. whom 27 were coinfected with human immunodeficiency virus type 1 (HIV-1). To overcome the technical problems in deThis uncertainty has rendered difficult both the understanding of HCV epidemiology and the sexual and reproductive counseltecting RNA in unfractionated semen and to understand which seminal fraction could serve as a reservoir for the virus, we ing of couples of whom only one partner, usually the male, is infected with HCV.separated the ejaculates into cell-free seminal plasma, seminal round cells, and spermatozoa before the PCR assay. To process The issue of sexual transmission of hepatitis viruses has become further complicated by the identification of a new semen we used a specific procedure that we have devised to treat HIV-infected semen [32]. HCV-related hepatitis virus named hepatitis G (HGV) [16] or 18]. It is now recognized that ...

Prevalence and genetic variants of hepatitis GB-C/HG and TT viruses in Gabon, equatorial Africa.

“…To avoid false positive results, we systematically introduced negative controls at each stage of PCR and dTTP was replaced by dUTP in the PCR mixture. 35 All positive and randomly selected negative samples gave the same result in two independent assays.…”

“…34 cDNA was synthesized by incubation at 42ЊC with MMLV reverse transcriptase and random hexamers (Perkin Elmer, Roche Molecular Systems, Inc., Branchburg, NJ) as previously described. 35 The cDNA was stored at Ϫ20ЊC until use.…”

“…For both PCR assays, amplification conditions were as previously described. 35 PCR products were assessed by Southern blot hybridization. 35 Detection of TTV DNA by PCR.…”

“…35 PCR products were assessed by Southern blot hybridization. 35 Detection of TTV DNA by PCR. DNA was extracted from 100 L serum using the Qiamp Blood kit (Qiagen S. A., Courtaboeuf, France) and dissolved in 100 L elution buffer.…”

Sexual activity as a risk factor for hepatitis C

“…3,4 Failure to detect HCV RNA in body secretions may be caused by technical factors, including specimen collection and storage, and the ability to exclude cellular components and to overcome the presence of polymerase chain reaction inhibitors. 4,5 Even in studies using optimal methods to isolate HCV RNA, the minority of samples were positive for HCV RNA and all positive samples were of low titer (equal to 10 2 copies/ mL). 4,6 A low titer of virus in genital secretions may be one reason that HCV is transmitted less efficiently than hepatitis B virus or HIV.…”