1998;55:203C208

1998;55:203C208. known. One hypothesis for the elevated prevalence of GBV-C markers in the blood donor population may be vertical transmission over several generations, similar to that of hepatitis B virus. Alternatively, GBV-C infections may be caused by sexual or even social contacts (8). The aim of the present study was to further characterize the transmission routes of GBV-C by analyzing for GBV-C markers in up to three generations of family members in three families with documented mother-to-infant transmission of GBV-C. Samples were collected from three previously identified families with known or suspected mother-to-infant transmission of GBV-C (4, 6). In family A, Thevetiaflavone where a male twin was vertically infected in 1993 (4), saliva and/or serum samples were obtained from both parents and the two twins in 1996, 1997, and 1999. In family B, where the child was suspected to have been infected by the mother between 1990 and 1993, serum samples were from the infected child (6), both parents, and the maternal grandparents in 1998. A stored sample from a more youthful brother acquired in 1993 was also analyzed. In family C, where vertical transmission occurred in 1991 (6), samples were from the infected son, the infected mother, and the maternal grandparents in 1998. All samples were analyzed for the presence of GBV-C RNA by a opposite transcription-PCR using primers from your 5-noncoding region as explained previously (2, 4). All serum samples were also tested at a dilution of 1 1:100 for antibodies Rabbit Polyclonal to ETS1 (phospho-Thr38) to a recombinant genotype 1 GBV-C E2 protein (kindly provided by I. K. Mushahwar, Abbott Laboratories) by an enzyme immunoassay (EIA) as previously explained (3). To confirm mother-to-infant transmission between the mother and the infected child in family B, the sequence of an 82-bp GBV-C nonstructural 3 fragment was identified and analyzed by phylogenetic analysis by standard protocols as previously explained (6). Cloning and manifestation of the E2 gene (mtE2) from your GBV-C genotype 2-infected male twin of family A (4) were performed in by standard techniques and will be explained in detail elsewhere. The mtE2 and the GBV-C E2 proteins were used in Western blot analysis for antibody detection after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PhastGelSystem; Pharmacia, Uppsala, Sweden) in Thevetiaflavone accordance with the manufacturer’s instructions. A hyperimmunized mouse serum (dilution 1:7,500) raised against the GBV-C E2 protein was used like a positive control in the Western blot. In family Thevetiaflavone A, only the previously explained mother and twin son (4) experienced serum GBV-C RNA in the 1997 and 1999 samples. Saliva samples from these two contained GBV-C RNA in 1996 (4) and 1997 but not in 1999 (Fig. ?(Fig.1).1). Neither of them experienced antibodies to GBV-C E2 or to the mtE2 protein as determined by EIA (Table ?(Table1)1) or by European blotting (Fig. ?(Fig.1).1). In contrast, both the father and the twin woman were bad for GBV-C RNA in serum but were positive for E2 antibodies in 1996 and 1997 as determined by EIA (Table ?(Table1)1) or by European blotting (Fig. ?(Fig.1),1), respectively. The twin woman developed antibodies to the mtE2 protein in 1996 and became clearly seropositive in 1997 as determined Thevetiaflavone by Western blotting (Fig. ?(Fig.1).1). However, since the antibodies did not identify the GBV-C E2 protein by EIA (data not demonstrated), we do not wish to overemphasize this observation. Her alanine aminotransferase and -glutamyl-transferase levels were persistently normal, whereas the asparagine aminotransferase levels.