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Objective: The aim of the study was to analyze the trends in HIV prevalence among childbearing women through unlinked anonymous testing of newborns. Methods: Dried-blood specimens, residual to routine metabolic screening of newborns, were collected in Italy during the last quarter of each year, from 1994 to 2002. These samples were tested anonymously for HIV antibodies. Results: Of 844,311 specimens tested, 703 (0.83/1000; 95% confidence interval, 0.76-0.88) resulted as HIV antibody-reactive. Prevalence was 1.05 per 1000 in 1994 and decreased significantly over the following 3 years, reaching 0.60 per 1000 in 1997. Thereafter, a seroprevalence increase was recorded with a figure of 1.01 per 1000 in 1999. A decrease was again observed in 2001 and in 2002, when the seroprevalence rate was significantly lower than that recorded in 1994. During the entire study period, higher prevalence rates were observed in northern Italian regions, where the highest AIDS incidence rates are recorded, and in metropolitan compared with nonmetropolitan areas. The estimated number of children born each year to an HIV-positive mother ranged from 313 to 546. Conclusion: The number of children born to an HIV-positive mother calls for continued efforts to prevent vertical transmission of the infection.

Assessment of the CD4 lymphocyte number, currently performed by flow cytometry, is one of the main laboratory tests for establishing progression to acquired immunodeficiency syndrome (AIDS). An enzyme immunoassay has recently been commercialized which can be very useful for counting CD4 cells in laboratories where flow cytometers are not available. In the present study, a comparative evaluation of CD4 positive lymphocytes with flow cytometry and the enzyme assay was made in healthy subjects (N = 30, R = 0.88, P < 0.0001), human immunodeficiency virus (HIV)-infected individuals (N = 80, R = 0.91, P < 0.0001), and patients with autoimmune diseases (N = 28, R = 0.82, P < 0.001) or psoriasis (N = 18, R = 0.76, P = 0.01). A correlation between the two methodologies was not found in psoriatic patients under treatment with cyclosporin A (N = 7, R = 0.05, not significant). Some differences could be found at low CD4 lymphocyte levels since the influence of CD4 antigen eluted from monocytes or soluble CD4 in the whole blood sample could cause overestimation of CD4 cell numbers by the enzyme assay. © 1995 Wiley-Liss, Inc.

IntroductionIn western countries the transmission of hepatitis B virus (HBV) transmission through multi-patients lancing devices has been inferred since early '90s, however no study has ever provided biological evidence which directly link these device with HBV cross-infection. Here we present results of an outbreak investigation which could associate, by molecular techniques, the use of lancing device on multiple patients with HBV transmission in an Italian oncohematology unit.MethodsThe outbreak investigation was designed as a retrospective cohort study to identify all potential cases. All cases identified were eventually confirmed through molecular epidemiology techniques. Audit of personnel including extensive review of infection control measures and reviewing personnel's tests for HBV was done identify transmission route.ResultsBetween 4 May 2006 and 21 February 2007, six incident cases of HBV infection were reported among 162 patients admitted in the oncohematology. The subsequent molecular instigation proved that 3 out 6 incident cases and one prevalent cases (already infected with HBV at the admission) represented a monophyletic cluster of infection. The eventual environmental investigation found that an identical HBV viral strain was present on a multi-patients lancing device in use in the unit and the inferential analysis showed a statistically significant association between undergoing lancing procedures and the infection.DiscussionThis investigation provide molecular evidence to link a HBV infection cluster to multi-patients lancing device and highlights that patients undergoing capillary blood sampling by non-disposable lancing device may face an unacceptable increased risk of HBV infection. Therefore we believe that multi-patients lancing devices should be banned from healthcare settings and replace with disposable safety lancets that permanently retract to prevent the use of the same device on multiple patients. The use of non-disposable lancing devices should be restricted to individual use at patients' home.

We describe a simple, sensitive, and reproducible method for using whole blood collected onto filter paper (dried blood spots) for detection and genotyping of hepatitis C virus RNA that can be useful in large field studies, particularly in settings where collection, preparation, storage, and shipment of samples at controlled temperature can be difficult.

Direct population sequencing and reverse hybridization (line probe assay [LiPA])-based methods are the most common methods for detecting hepatitis B virus (HBV) drug resistance mutations, although only mutations present in viral quasispecies with a prevalence of ≥20% can be detected by sequencing, and only known mutations are detected by LiPA. Massively parallel ultradeep pyrosequencing (UDPS; GS FLX platform) was used to analyze HBV quasispecies in reverse transcriptase (RT) and hepatitis B S antigen (HBsAg) from five drug-naive patients and eight drug-resistant patients. Eight primer pairs were used to obtain partially overlapping amplicons, covering the RT gene from codons 1 to 288 and the complete overlapping HBsAg sequence. A 1% mutation frequency was selected as the cutoff based on an error rate estimated on plasmid DNA. This technology enabled simultaneous analysis of between 2,852 and 18,016 clonally amplified fragments from each patient. The results indicate that UDPS has a relative sensitivity much higher than both direct sequencing and LiPA. In addition, the UDPS results are quantitative, allowing establishment of the relative frequency of both known mutations and novel substitutions. Some of the detected RT substitutions led to changes also in HBsAg. On the whole, genotype D presented a higher heterogeneity than genotype A. Considering the high quantity of information that can be provided by a single test from one patient, the short turnaround time, the information on substitution frequency, and the detection of rare variants, there are strong advantages conferred by UDPS, and the new method could play a relevant role in the clinical management of HBV infection and therapy.

Intra-hepatic levels of mRNA for IFN-a, IFN-y, IFN type 1 receptor (IFNAR-1) and PKR were determined in hepatitis C virus (HCV)/HIV co-infected and HCV mono-infected patients. In co-infected patients, IFN-a mRNA was upregulated and correlated with HIV-1 viraemia. IFN-y, IF-NAR-1 and PKR mRNA were detected in mono-infected, but not in co-infected patients. These findings suggest that in HCV/HIV co-infected individuals the ability to respond to IFN-a is impaired, probably because of the absence of its receptor.

HCV and interferon: viral strategies for evading innate defence mechanisms in the virus-host battle

Concerns about safety of rituximab in hepatitis B virus (HBV) carriers have risen at the light of reported cases of HBV reactivation following the administration of this agent. We recently observed a patient affected by B-cell Chronic Lymphocytic Leukemia (B-CLL) who developed this complication without any serological evidence of HBsAg expression. Case report: the patient, a 51 old year’s male, was diagnosed as having B-CLL in November 1998 and followed until his death due to acute liver failure due to HBV reactivation of a mutant HBV strain in September 2004 (Table). At diagnosis of B-CLL no risk factors regarding liver diseases or viral infections were reported; serum HBsAg and HBeAg, antibodies against the B-core antigen (HBcAb), HBe (HBeAB), HBsAg (HBsAb), HCV, hepatitis A and Delta virus, Cytomegalovirus, and Epstein Barr virus were negative while HBcAb alone were positive. He presented stable disease until November 2001, when fludarabine was started because of progressive disease. A good partial response (GPR) was achieved so that he received four weekly standard doses (375 mg/m2) of rituximab as consolidation therapy (July 2002). A complete remission (CR) was obtained and he was then closely followed-up until about two years later (February 2004) when, for a disease recurrence, rituximab was given again. At that time all the above hepatitis markers remained unmodified. After four weekly standard doses of rituximab a GPR was recorded, and then six monthly courses (150 mg/m2) of the same agent were administered as maintenance therapy. In September 2004, being the patient in CR, he was admitted with acute hepatitis. HBV DNA strains (200,000 copies/mL) and HbcAb IgM were detected, whereas HbsAg-negative status persisted. The viral form was characterized as having D genotype and ayw3 serotype respectively. Sequence analysis of polymerase chain reaction (PCR) products amplified from the S region revealed 5 remarkable mutations in the major antigenic regions (C124Y, A128V, G130D, P135R and G145R). These mutations were associated with the lack of HbsAg production and were compatible with an escape of a rare HBV strain from the patient’s own HbsAb. Despite treatment with lamivudine, the patient died of fulminant hepatic failure. Conclusion: our experience indicates that patients receiving rituximab who are negative for HBsAg but positive for anti-HBc are still at risk for reactivation of latent HBV and should be considered for HBV DNA testing and prophylaxis with lamivudine. Patient clinical features Clinical features Diagnosis (November 1998) Disease Progression (November 2001) Disease Recurrence (February 2004) B Hepatitis onset (September 2004) *D genotype, ayw3 serotype; C124Y, A128V, G130D, P135R and G145R mutations in the S region. NA:not available. Rai Clinical Stage I II II 0 CD38 Negative Negative Negative Negative Zap 70 Positive Positive Positive Negative CD4/CD8 0.95 0.90 0.85 0.80 Beta 2 microglobulin (mcg/L) 1503 1825 1844 1325 Serum Ig level (gr/L) 0,980 0,790 0,660 0,620 HBsAg Negative Negative Negative Negative HBsAb Negative Negative Negative Negative HBcAb Positive Positive Positive Positive HBeAg Negatrive Negative Negative Positive HBeAb Positive Positive Positive Positive HBV DNA* NA NA NA 200.000/μL