Prevalence and follow-up of occult HCV infection in an Italian population free of clinically detectable infectious liver disease.

BACKGROUND: Occult hepatitis C virus infection (OCI) is a recently described phenomenon characterized by undetectable levels of HCV-RNA in serum/plasma by current laboratory assays, with identifiable levels in peripheral blood mononuclear cells (PBMCs) and/or liver tissue by molecular tests with enhanced sensitivity. Previous results from our group showed an OCI prevalence of 3.3% in a population unselected for hepatic disease. The present study aimed to evaluate OCI prevalence in a larger cohort of infectious liver disease-free (ILDF) subjects. Clinical follow-up of OCI subjects was performed to investigate the natural history of the infection. METHODS AND FINDINGS: 439 subjects referred to a Turin Blood Bank for phlebotomy therapy were recruited. They included 314 ILDF subjects, 40 HCV-positive subjects and 85 HBV-positive subjects, of whom 7 were active HBV carriers. Six subjects (4/314 ILDF subjects [1.27%] and 2/7 active HBV carriers [28%]) were positive for HCV-RNA in PBMCs, but negative for serological and virological markers of HCV, indicating OCI. HCV genotypes were determined in the PBMCs of 3/6 OCI subjects two had type 1b; the other had type 2a/2c. OCI subjects were followed up for at least 2 years. After 12 months only one OCI persisted, showing a low HCV viral load (3.73×10(1) UI/ml). By the end of follow-up all OCI subjects were negative for HCV. No seroconversion, alteration of liver enzyme levels, or reduction of liver synthesis occurred during follow-up. CONCLUSIONS: This study demonstrated the existence of OCI in ILDF subjects, and suggested a high OCI prevalence among active HBV carriers. Follow-up suggested that OCI could be transient, with a trend toward the decrease of HCV viral load to levels undetectable by conventional methods after 12-18 months. Confirmation studies with a longer follow-up period are needed for identification of the OCI clearance or recurrence rates, and to characterize the viruses involved.

Thiopurine S-methyltransferase pharmacogenetics in a large-scale healthy Italian-Caucasian population: differences in enzyme activity.

AIMS: To investigate the influence of genotype, age and gender on the thiopurine S-methyltransferase (TPMT) phenotype in healthy Italian-Caucasian subjects. MATERIALS & METHODS: The study investigated the TPMT genotype and the TPMT phenotype of 943 healthy Italian-Caucasian subjects of different age and gender (age range: 0.08-68 years; 623 males 320 females). TPMT red blood cell activity was measured in all samples and genotype was determined for the TPMT alleles *2, *3A, *3B and *3C. RESULTS: TPMT activity levels in our whole population ranged from 1.6 up to 75.2 U/gHb. Significant TPMT activity differences between wild-type and heterozygous subjects were observed. We divided our TPMT activity into four categories according to our frequency distribution: low (0.1%), intermediate (32.9%), normal (60%) and high (7%), with arbitrary cut-off values of 8.0, 19.4 and 37.0 U/gHb, respectively. The whole population had a total of 94.5% of homozygous wild-type subjects, 5.4% heterozygous variants and one (0.1%) compound heterozygous variant TPMT*3B/*3C. The overall concordance rate between TPMT genotypes and phenotypes was 71.6%. The TPMT activity was significantly higher in wild-type children (0.08-17 years) than in wild-type adults (aged 18-68 years). Moreover, it was noted that wild-type infants from 0.08 to 5 years had a 9% higher average TPMT activity than the other wild-type groups, and only in children from 0.08 to 2 years was the TPMT activity higher in males than in females. CONCLUSION: The data obtained in this study show that genetic factors seem to be the major aspect in TPMT phenotype variability in adults, whilst, in children, other physiological factors should be taken into consideration when assessing the TPMT phenotype, such as age and gender.

Evidence of acute primary occult hepatitis B virus infection in an Italian repeat blood donor.

BACKGROUND: Preliminary evidence of cases of acute occult hepatitis B virus (HBV) infection (OBI) has been recently reported in the literature. Furthermore, OBI definition has been the object of an international consensus conference. STUDY DESIGN AND METHODS: A case of acute primary OBI was identified and followed up in a repeat female blood donor using a highly sensitive nucleic acid test (NAT; Procleix Ultrio on Tigris, Chiron). Genotyping and sequencing of virus isolates from donor and contact cases were performed. RESULTS: The blood donor never developed detectable hepatitis B surface antigen (HBsAg) until seroconversion to antibody to hepatitis B surface antigen/antibody to hepatitis B core antigen. A very low viral load was observed during the infection course (<50 IU/mL). Donor HBV DNA sequencing consistently showed a CCA deletion leading to amino acid T116 deletion in the small envelope protein (S). Other sequence features showed high homology between donor and contact case, suggesting a sexual transmission. DISCUSSION: The main explanation for HBsAg undetectability relies on the very low level of viremia observed. The single-amino-acid deletion found in the S protein cannot account for HBsAg detection failure, because the capture antibody of the assay used is targeted to a different sequence epitope (aa121-124). Meanwhile, CCA deletion may have impacted the virus replication efficiency since it affects the overlapping reverse transcriptase "finger" domain of the polymerase gene. These findings define this case as an acute primary OBI, confirming the existence of this condition. NAT with high sensitivity is the only screening enabling prevention of HBV transmission by transfusion in such cases.

Italian blood donors with anti-HBc and occult hepatitis B virus infection.

BACKGROUND AND OBJECTIVES: Occult hepatitis B virus (HBV) infection might allow the release of viremic units into the blood supply network if blood is tested only for hepatitis B surface antigen (HBsAg). The aim of our study was to evaluate the actual prevalence, viral load and genotype of occult HBV infections among first-time blood donors in north-western Italy and to suggest a way to minimize risks of transmission of this infection. DESIGN AND METHODS: We assayed 6313 consecutive blood donors for antibodies to HBV core antigen (anti-HBc) in addition to mandatory screening. HBsAg-negative/anti-HBc-positive donors were assayed for antibodies to HBsAg (anti-HBs) and for HBV-DNA using COBAS Ampliscreen HBV (Roche) on individual donations. All HBV-DNA-positive samples underwent confirmatory testing with additional polymerase chain reaction-based assays. RESULTS: The prevalence of anti-HBc positive subjects was 4.85%. Fourteen out of 288 blood donors (4.86%) were confirmed to have circulating HBV-DNA at a low level (range 8-108 IU/mL). All viremic donors were also anti-HBs-positive. INTERPRETATION AND CONCLUSIONS: We estimate that in north-western Italy up to 2298 units per million donated units from first-time donors may contain HBV-DNA. The risk of an HBV-DNA positive unit from an occult carrier being released into the blood supply is more than 100 times higher than the estimated residual risk related to the window phase of HBV infection in our country. The potential infectivity of these units is debated, but their use cannot be considered safe at least in immunocompromised patients.

Soliton-based Y-branch in photorefractive crystals induced through dispersion-shifted optical fiber.

We experimentally demonstrate for what is believed to be the first time that a dispersion-shifted fiber can be used to electro-optically induce a soliton Y-branch structure in a photorefractive centrosymmetric paraelectric crystal (potassium lithium tantalate niobate). The application of a nonstationary external bias field enables us to stabilize the spatially partially coherent behavior of the optical beam at the fiber output. Furthermore, we show the switching capabilities of this soliton-based device in the optical communication field guiding a probe beam at a nonphotorefractive wavelength (1557 nm).