To determine the distribution of hepatitis C virus (HCV) genotypes and to related genotype to viral load, genotyping and quantification of viral RNA were carried out in 35 patients with chronic hepatitis C. Subtype 1a was most prevalent (43%), followed by subtypes 1b (23%) and 3a (14%). Mean viral load (log HCV-RNA copies/ml) for subtypes 1b, 1a and 3a was 7.1 +/- 1, 5.6 +/- 1.1 and 4.1 +/- 2.4, respectively. The presence of immunoglobulin M was related to the duration of hepatitis and genotype 1 to a more severe hepatic injury and higher viral load. Differences observed in viral load for a single HCV subtype justify the need to quantify HCV-RNA prior to establishing antiviral therapy.

Helicobacter pylori is associated with peptic ulcer and chronic active gastritis. The response to infection can be determined by measuring serum titers of anti-H. pylori antibodies. We compared antibody titers in 612 serum samples from 570 individuals considered at risk for H. pylori infection, 170 of them are control sera from 110 adults and 60 children with no gastric alterations. The study groups were 93 institutionalized mentally handicapped children, 40 heterosexual couples, 101 HIV-sero-positive patients, 86 patients with chronic renal failure and 40 individuals (20 adults and 20 children) with symptoms associated with gastritis or gastroduodenal ulcer disease. In the adult and child control groups, 33.5% and 11.6% of the individuals had circulating anti-H. pylori antibodies. Significantly more adults (80%) and children (75%) with gastric symptoms had detectable circulating antibody titers. Elevated titers were also found in institutionalized children and in adults with renal failure.

SUMMARYIntroduction. Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests.Methods. After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen.Results. In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions.Conclusions. Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

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We have evaluated a new enzyme immunoassay technology to detect the products of PCR-based amplification that may be applicable to routine testing of hepatitis B virus (HBV) DNA. Two hundred eight serum samples were studied: 73 were basal samples and 135 were sequential serum samples from patients with chronic hepatitis, some of whom were being treated with alpha interferon. We compared the new detection method (PCR-DNA enzyme immunoassay [DEIA]) with dot blot hybridization performed without prior PCR amplification and with two other methods for detection of PCR products: agarose gel electrophoresis with ethidium bromide staining (PCR-EB) and dot blot (PCR-dot blot). For hepatitis B-antigen-positive basal samples, HBV DNA was detected in 70.4% by dot blot, 74.1% by PCR-EB, and 100% by PCR-DEIA and PCR-dot blot; for anti-hepatitis B e-antigen basal samples, HBV DNA was found in 10.5% by dot blot and PCR-EB and in 42.1% by PCR-DEIA and PCR-dot blot. Chi-square tests showed a strong association between dot blot and PCR-EB and between PCR-DEIA and PCR dot blot. Using PCR-dot blot as the reference, dot blot shows a 56.9% sensitivity and a 100% specificity, PCR-EB shows a 55.0% sensitivity and a 100% specificity, and PCR-DEIA shows a 95.4% sensitivity and a 97% specificity. We conclude that the technical advantages of the DEIA method and its high sensitivity and specificity may facilitate the use of PCR in routine testing for HBV DNA in clinical microbiology laboratories.