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ABSTRACT: Human papillomavirus (HPV) has been identified as the major cause of cervical cancer worldwide and HPV DNA testing is recommended in primary cervical cancer screening. Several molecular tests for detection/typing of HPV DNA with different sensitivity and specificity are commercially available. The present study compared the performance of the Abbott RealTime High Risk HPV assay and the Genomica HPV Clinical Array CLART2 in 78 specimens (63 cervical smears and 15 rectal/urethral swabs).The typing results of the Genomica assay were in absolute agreement with each of the four possible result categories of the Abbott assay (HPV16, HPV18, Other HR HPV, not detected) in 87.2% (68/78) of the samples, with a Cohen' kappa agreement coefficient for every HR type of 0.62 (95% CI: 0.39-0.85), higher in cervical swabs (k = 0.74, 95% CI: 0.50-0.99) than in rectal/urethral swabs (k = 0.36, 95% CI: 0.00-0.82). There was an excellent agreement of the Genomica results with those of Abbott in cervical samples harbored HPV single infection (100% agreement). Nonetheless, both methods may lose sensitivity for detecting HPV types in multiple infections, giving discordant results (10/78). This underlines the importance of establishing the analytical sensitivity in HPV type detection in single and multiple HPV infections. In rectal/urethral swabs, 5 of 15 (33%) discordant cases were observed, most of which became compatible when the Genomica assay was performed starting from nucleic acid extracted with the Abbott m2000sp system. These results suggest that nucleic extraction based on the magnetic beads technique is suitable for HPV DNA detection in urethral/rectal swabs.
Apmis 02/2013; · 1.99 Impact Factor
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Valentina Svicher,
Valeria Cento,
Gabriella Rozera,
Isabella Abbate,
Maria Mercedes Santoro,
Daniele Armenia,
Lavinia Fabeni,
Alessandro Bruselles,
Alessandra Latini,
Guido Palamara,
Valeria Micheli,
Giuliano Rizzardini,
Caterina Gori,
Federica Forbici,
Giuseppe Ippolito,
Massimo Andreoni,
Andrea Antinori,
Francesca Ceccherini-Silberstein, Maria Rosaria Capobianchi,
Carlo Federico Perno
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ABSTRACT: The false-positive rate (FPR) is a percentage-score provided by Geno2Pheno-algorithm indicating the likelihood that a V3-sequence is falsely predicted as CXCR4-using. We evaluated the correlation between FPR obtained by V3 population-sequencing and the burden of CXCR4-using variants detected by V3 ultra-deep sequencing (UDPS) and Enhanced-Sensitivity Trofile assay (ESTA).
54 HIV-1 B-subtype infected-patients (all maraviroc-naïve), with viremia >10,000copies/ml, were analyzed. HIV-tropism was assessed by V3 population-sequencing, UDPS (considering variants with >0.5% prevalence), and ESTA.
By UDPS, CCR5-using variants were detected in 53/54 patients, irrespective of FPR values, and their intra-patient prevalence progressively increased by increasing the FPR obtained by V3 population-sequencing (rho = 0.75, p = 5.0e-8). Conversely, the intra-patient prevalence of CXCR4-using variants in the 54 patients analyzed progressively decreased by increasing the FPR (rho = -0.61; p = 9.3e-6). Indeed, no CXCR4-using variants were detected in 13/13 patients with FPR>60. They were present in 7/18 (38.8%) patients with FPR 20-60 (intra-patient prevalence range: 2.1%-18.4%), in 5/7 (71.4%) with FPR 10-20, in 4/6 (66.7%) with FPR 5-10, and in 10/10(100%) with FPR<5 (intra-patient prevalence range: 12.1%-98.1%).
FPR by V3 population-sequencing can predict the burden of CXCR4-using variants. This information can be used to optimize the management of tropism determination in clinical practice. Due to its low cost and short turnaround time, V3 population-sequencing may represent the most feasible test for HIV-1 tropism determination. More sensitive methodologies (as UDPS) might be useful when V3 population-sequencing provides a FPR >20 (particularly in the range 20-60), allowing a more careful identification of patients harboring CXCR4-using variants.
PLoS ONE 01/2013; 8(1):e53603. · 4.09 Impact Factor
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Caterina Mele,
Paraskevas Iatropoulos,
Roberta Donadelli,
Andrea Calabria,
Ramona Maranta,
Paola Cassis,
Simona Buelli,
Susanna Tomasoni,
Rossella Piras,
Mira Krendel, [......],
Friedhelm Hildebrandt,
Edgar Otto,
Franz Schaefer,
Fabio Macciardi,
Fatih Ozaltin,
Sevinc Emre,
Tulin Ibsirlioglu,
Ariela Benigni,
Giuseppe Remuzzi,
Marina Noris
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Giada Rossini,
Fabrizio Carletti,
Roberto Rigoli,
Sandro Piga,
Patrizia Bagnarelli,
Paolo Gaibani,
Anna Pierro,
Alessandro Nanni Costa,
Paolo Grossi,
Giuseppe Ippolito,
Maria Paola Landini,
Antonino Di Caro, Maria Rosaria Capobianchi,
Vittorio Sambri
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ABSTRACT: West Nile virus (WNV) is currently circulating in several European countries and over the past decades, concomitant with enhanced surveillance studies and improved diagnostic capabilities, an increase in the geographical distribution and in the number of cases in Europe has been documented. In Italy, in 2011, 14 human cases of WNV neuroinvasive infections due to lineage 1 strains were registered in several Italian regions. Here we report WNV partial sequences obtained from serum samples of two patients living in different regions in Italy (Veneto and Sardinia). Phylogenetic analysis, performed on a fragment (566 nt) of the Envelope gene, showed that WNV strains circulating in Italy in 2011 belong to lineage 1a but are different from lineage 1a strains isolated in 2008-2009.The data here reported are consistent with the hypothesis of multiple recent introductions in Italy of WNV lineage 1a strains.
Journal of General Virology 11/2012; · 3.36 Impact Factor
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ABSTRACT: The management of viral hemorrhagic fevers (VHFs) has mainly focused on strict infection control measures, while standard clinical interventions that are provided to patients with other life-threatening conditions are rarely offered to patients with VHFs. Despite its complexity, a proper clinical case management of VHFs is neither futile nor is it lacking in scientific rationale. Given that patient outcomes improve when treatment is started as soon as possible, development and implementation of protocols to promptly identify and treat patients in the earliest phases of diseases are urgently needed. Different pharmacological options have been proposed to manage patients and, as for other life-threatening conditions, advanced life support has been proved effective to address multiorgan failure. In addition, high throughput screening of small molecular libraries has emerged as a novel promising way to find new candidates drugs for VHFs therapy and a relevant number of new molecules are currently under investigation. Here we discuss the current knowledge about VHF clinical management to propose a way to step up the approach to VHFs beyond the mere application of infection control measures.
BMC Medicine 03/2012; 10:31. · 6.03 Impact Factor
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Chiara Agrati,
Cristiana Gioia,
Concetta Castilletti,
Daniele Lapa,
Giulia Berno,
Vincenzo Puro,
Fabrizio Carletti,
Eleonora Cimini,
Carla Nisii,
Flora Castellino,
Federico Martini, Maria R Capobianchi
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ABSTRACT: Abstract Influenza vaccination is recommended for HAART-treated HIV patients to prevent influenza illness and complications. Due to the known ability of T cells to mediate a broadly cross-reactive response, vaccination effectiveness in cell-mediated immune (CMI) response induction is a main objective in new influenza vaccination strategies. Nevertheless, data on CMI responses after pandemic vaccination in HIV subjects are still missing. In the present study, the ability of a single dose of adjuvanted pandemic influenza vaccine to induce humoral and CMI responses was compared in HAART-treated HIV patients and in healthcare workers. Healthcare workers (HCW, n=65) and HAART-treated HIV patients (HIV, n=67) receiving pandemic vaccination were enrolled and analyzed before (t0) and after (t1) vaccination. The analysis of strain-specific humoral response was performed by HAI assay; CMI against pandemic (A/H1N1/Cal/09) and seasonal (A/H1N1/Brisb/07 and A/H3N2/Brisb/07) strains was analyzed by ELISpot and intracellular staining followed by flow cytometry. Pandemic vaccination was effective in inducing both humoral and cell-mediated responses in HAART-treated HIV patients as well as in HCWs. A large fraction of both HCWs and HIV-infected patients showed a T cell response to the pandemic strain before vaccination, suggesting possible previous exposure to A/H1N1/pdm/09 and/or cross-reactive T cells. Notably, pandemic vaccine was also able to boost cross-reactive immune responses to seasonal strains. Finally, a weaker boost of both strain-specific and cross-reactive T cell immunity was found in individuals showing a higher baseline response. These data show the effectiveness of adjuvanted pandemic vaccine to induce both humoral and cellular (strain-specific and cross-reactive) immune responses in HIV patients similar to HCWs.
AIDS research and human retroviruses 03/2012; · 2.18 Impact Factor
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ABSTRACT: Natural history of HPV infection is altered in HIV positive women, with increased risk of cervical dysplasia. Limited data are available about the influence of HPV genotypes and HPV multiple infections on cervical disease in HIV positive women.
We determined HPV genotype prevalence in cervical samples from 553 HIV-infected women attending the gynaecological service at "L. Spallanzani" Hospital. Association of HPV multiple infections with cervical abnormalities was investigated.
MY09/MY11 consensus primers were used to detect HPV-DNA; HPV typing was performed by RFLP. A commercial array based kit was used to define unresolved RFLP patterns.
HPV was detected in 244/553 (44.1%) patients, correlating with low CD4 counts (p<0.001) and age (p=0.001). Multiple HPV types were observed in 44.4% of cases, more frequently involving HR than LR HPV (OR=12.8, p<0.00001). Multiple HPV infections were associated with low CD4 counts (OR=3.8 in CD4<200 vs CD4≥500 cells/mm(3)). Dyskaryosis was associated with decreased CD4 counts (≥500 vs 200-499 vs <200 cells/mm(3), χ(2) for trend, p=0.001) and with HPV types multiplicity (1 vs 2-3 vs ≥4, χ(2) for trend, p<0.00001). Notably, in 3 H-SIL cases only LR types were detected (HPV62, n=2; HPV81, n=1).
Multiple HPV infections, often involving HR types, are frequent in HIV-infected women. Association between multiple HPV infection, low CD4 count and cytological abnormalities supports the interplay of virological and immunological factors in cervical cancer pathogenesis. Assessment of multiple HPV infections might gain importance in cervical cancer screening, particularly in patients with predisposing factors like immuno-suppression.
Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 03/2012; 54(2):141-6. · 3.12 Impact Factor
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ABSTRACT: Outbreaks of Acute Hepatitis A Virus (HAV) among men who have sex with men (MSM) have been reported in Europe and, recently, in Italy. From July 2008 through January 2010, 162 HAV infections were diagnosed at National Institute for Infectious Diseases, Rome, Italy, with high male-to-female ratio (M:F=7.5).
The aim of this study was to characterize viral strains involved in this outbreak.
The sequences of VP1-2A junction of HAV genome, obtained from 67/97 HAV-RNA-positive samples, were used for phylogenetic analysis.
All but 1 of the HAV sequences were genotype 1A, 1 was genotype 1B. A monophyletic cluster, including 59/66 genotype IA sequences, was identified by phylogenetic analysis. This cluster included also 2 HAV strains isolated in Germany (2007) and France (2008) from MSM, that, in turn, were reported to be genetically correlated to HAV strains circulating in Tuscany in 2008. Among the males harboring an HAV strain belonging to the cluster, 62% reported to be MSM, and 25% were HIV-positive, 2 with acute HIV infection.
The outbreak occurred in Rome in 2008-2010, involving high proportion of HIV-infected MSM, is sustained by a monophyletic HAV strain, circulating around the same period also in other European countries. Possible factors favouring HAV spread among HIV-infected persons, such as high risk behavior and prolonged fecal excretion, need to be further elucidated. Timely identification of outbreaks with one or the same source of infection may be helpful to implement preventive measures addressing at risk populations.
Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 02/2012; 54(1):26-9. · 3.12 Impact Factor
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Silvia Nozza,
Filippo Canducci,
Laura Galli,
Alessandro Cozzi-Lepri, Maria Rosaria Capobianchi,
Elisa Rita Ceresola,
Pasquale Narciso,
Raffaella Libertone,
Paula Castelli,
Mariacristina Moioli,
Antonella D'Arminio Monforte,
Antonella Castagna
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ABSTRACT: To investigate the value of tropism (determined by genotypic testing) to predict CD4 depletion in HIV-infected antiretroviral-naive patients with high CD4 counts.
Viral tropism was determined by geno2pheno (false positive rate = 10%) in 223 HIV-infected subjects naive to antiretrovirals with CD4 count ≥350 cells/μL and HIV-RNA >500 copies/mL enrolled in the ICONA Foundation Study for whom a stored plasma sample (baseline) was retrospectively tested. We monitored CD4 cell count and identified predictors of decline before antiretroviral therapy initiation, applying a mixed linear model with covariates (age, gender, tropism, HIV risk factor, calendar year of HIV infection, months from HIV diagnosis to baseline, hepatitis C virus status, CD4 and HIV-RNA at sample collection and duration of follow-up).
Two hundred and twenty-three subjects met the eligibility criteria; 137 (61%) were male and the median age was 35 (31-40) years. Median follow-up was 16.4 (3.2-37.2) months. Median CD4 decrease during follow-up was -157 (-278 to -13) cells/μL. At baseline, 192 (86%) subjects were defined as harbouring R5 virus and 31 (14%) non-R5. Median CD4 count was 571 (458-729) cells/μL and median HIV-RNA was 4.08 (3.57-4.55) log(10) copies/mL. At multivariable analysis, a greater mean CD4 decrease was associated with non-R5 viral tropism (-159.9 ± 12.22, P = 0.0002) at baseline. Other significant covariates were female gender, older age, intravenous drug use, longer duration of follow-up, and higher CD4 cell count and higher HIV-RNA at sample collection.
In patients with CD4 counts ≥350 cells/μL, non-R5 viral tropism by geno2pheno is predictive of CD4 decrease independent of their viral set point and CD4 counts.
Journal of Antimicrobial Chemotherapy 01/2012; 67(5):1224-7. · 5.07 Impact Factor
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Eleonora Cimini,
Cécile Bonnafous,
Veronica Bordoni,
Eleonora Lalle,
Helene Sicard,
Alessandra Sacchi,
Giulia Berno,
Cristiana Gioia,
Gianpiero D'Offizi,
Ubaldo Visco Comandini,
Chrysoula Vlassi, Maria Rosaria Capobianchi,
Federico Martini,
Chiara Agrati
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ABSTRACT: In chronic HCV infection, treatment failure and defective host immune response highly demand improved therapy strategies. Vγ9Vδ2 T-cells may inhibit HCV replication in vitro through IFN-γ release after Phosphoantigen (PhAg) stimulation. The aim of our work was to analyze Vγ9Vδ2 T-cell functionality during chronic HCV infection, studying the role of IFN-α on their function capability. IFN-γ production by Vγ9Vδ2 T-cells was analyzed in vitro in 24 HCV-infected patients and 35 healthy donors (HD) after PhAg stimulation with or without IFN-α. The effect of in vivo PhAg/IFN-α administration on plasma IFN-γ levels was analyzed in M. fascicularis monkeys. A quantitative analysis of IFN-γ mRNA level and stability in Vγ9Vδ2 T-cells was also evaluated. During chronic HCV infection, Vγ9Vδ2 T-cells showed an effector/activated phenotype and were significantly impaired in IFN-γ production. Interestingly, IFN-α was able to improve their IFN-γ response to PhAg both in vitro in HD and HCV-infected patients, and in vivo in Macaca fascicularis primates. Finally, IFN-α increased IFN-γ-mRNA transcription and stability in PhAg-activated Vγ9Vδ2 T-cells. Altogether our results show a functional impairment of Vγ9Vδ2 T-cells during chronic HCV infection that can be partially restored by using IFN-α. A study aimed to evaluate the antiviral impact of PhAg/IFN-α combination may provide new insight in designing possible combined strategies to improve HCV infection treatment outcome.
PLoS ONE 01/2012; 7(5):e37014. · 4.09 Impact Factor
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Francesco Vairo,
Emanuele Nicastri,
Silvia Meschi,
Monica Sanè Schepisi,
Maria Grazia Paglia,
Nazario Bevilacqua,
Sabina Mangi,
Maria Rosaria Sciarrone,
Roberta Chiappini,
Jape Mohamed,
Vincenzo Racalbuto,
Antonino Di Caro, Maria Rosaria Capobianchi,
Giuseppe Ippolito
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ABSTRACT: Evidence available to date indicates that dengue viruses 1, 2, and 3 could be among the causes of acute fever in eastern Africa. Recently, four reports on dengue infection in travelers and residents have raised concerns over the occurrence of dengue fever in mainland Tanzania and in Zanzibar. The objective of this study was to provide seroprevalence data on dengue infection in Tanzania.
This study was conducted in 2007 at two peripheral hospitals, one on Pemba Island, Zanzibar and one in Tosamaganga, Iringa Region, mainland Tanzania. Two hundred and two consecutive febrile outpatients were studied for antibodies and viral RNA to assess the circulation of dengue virus in Tanzania.
A seroprevalence of 7.7% was found on Pemba Island and of 1.8% was found in Tosamaganga. No acute cases and no previous infections among patients under 11 years of age were detected.
These findings provide the first baseline data on dengue seroprevalence in the country. No recent dengue virus circulation in Tanzania and in the Zanzibar archipelago up until the early 1990s is reported.
International journal of infectious diseases: IJID: official publication of the International Society for Infectious Diseases 11/2011; 16(1):e44-6. · 2.17 Impact Factor
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Claudia Alteri,
Maria Mercedes Santoro,
Isabella Abbate,
Gabriella Rozera,
Alessandro Bruselles,
Barbara Bartolini,
Caterina Gori,
Federica Forbici,
Nicoletta Orchi,
Valerio Tozzi,
Guido Palamara,
Andrea Antinori,
Pasquale Narciso,
Enrico Girardi,
Valentina Svicher,
Francesca Ceccherini-Silberstein, Maria Rosaria Capobianchi,
Carlo Federico Perno
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ABSTRACT: This proof-of-concept study aimed to identify whether mutations considered not yet relevant for drug resistance (but located at key drug-resistance positions) can act as 'sentinels' of minority resistant variants in HIV-1 drug-naive patients.
We focused our attention on three reverse transcriptase (RT) mutations (T69S, L210M and K103R) easily detected by standard population sequencing [i.e. the genotypic resistance test (GRT)]. Ultra-deep pyrosequencing (UDPS) of HIV-1 RT was performed using GS-FLX Roche, on plasma RNA from 40 drug-naive patients infected with HIV-1 subtype B without primary resistance detected by GRT. Only RT drug resistance mutations detected at >0.1% in both forward and reverse directions were considered. Associations between GRT sentinel mutations and UDPS drug resistance were assessed using Fisher's exact test.
UDPS detected drug resistance mutations in 18/40 drug-naive patients. Patients carrying HIV-1 strains with T69S and L210M by GRT showed a trend to greater infection by minority drug-resistant variants than control patients infected by HIV-1 without these mutations (5/10 and 7/10 versus 3/10; P = not significant). No association was found for K103R by GRT. Notably, T69S and L210M (but not K103R or control viruses) were associated with GRT minority drug-resistant variants with a prevalence >1% (3/10 and 4/10 versus 0/20 in K103R and controls; P = 0.03 and P = 0.008, respectively). Moreover, the presence of L210M or T69S viruses by GRT significantly correlated with that of minority thymidine analogue mutations by UDPS (6/20 patients carrying HIV-1 strains with T69S/L210M versus 0/20 patients carrying HIV-1 having K103R or none of these mutations; P = 0.03).
This proof-of-concept study suggests the existence of genetic markers, detectable by routine testing, potentially acting as sentinel mutations of minority drug resistance. Their identification may help in the selection of patients at high risk of resistance in reservoirs without the necessity of using UDPS.
Journal of Antimicrobial Chemotherapy 09/2011; 66(11):2615-23. · 5.07 Impact Factor
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ABSTRACT: Numerous studies investigating clinical significance of HIV-1 minimal residual viremia (MRV) suggest potential utility of assays more sensitive than those routinely used to monitor viral suppression. However currently available methods, based on different technologies, show great variation in detection limit and input plasma volume, and generally suffer from lack of standardization.
In order to establish new tools suitable for routine quantification of minimal residual viremia in patients under virological suppression, some modifications were introduced into standard procedure of the Abbott RealTime HIV-1 assay leading to a "modified" and an "ultrasensitive" protocols.
The following modifications were introduced: calibration curve extended towards low HIV-1 RNA concentration; 4 fold increased sample volume by concentrating starting material; reduced volume of internal control; adoption of "open-mode" software for quantification. Analytical performances were evaluated using the HIV-1 RNA Working Reagent 1 for NAT assays (NIBSC). Both tests were applied to clinical samples from virologically suppressed patients.
The "modified" and the "ultrasensitive" configurations of the assay reached a limit of detection of 18.8 (95% CI: 11.1-51.0 cp/mL) and 4.8 cp/mL (95% CI: 2.6-9.1 cp/mL), respectively, with high precision and accuracy. In clinical samples from virologically suppressed patients, "modified" and "ultrasensitive" protocols allowed to detect and quantify HIV RNA in 12.7% and 46.6%, respectively, of samples resulted "not-detectable", and in 70.0% and 69.5%, respectively, of samples "detected <40 cp/mL" in the standard assay.
The "modified" and "ultrasensitive" assays are precise and accurate, and easily adoptable in routine diagnostic laboratories for measuring MRV.
Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 09/2011; 52(1):17-22. · 3.12 Impact Factor
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Caterina Mele,
Paraskevas Iatropoulos,
Roberta Donadelli,
Andrea Calabria,
Ramona Maranta,
Paola Cassis,
Simona Buelli,
Susanna Tomasoni,
Rossella Piras,
Mira Krendel, [......],
Friedhelm Hildebrandt,
Edgar Otto,
Franz Schaefer,
Fabio Macciardi,
Fatih Ozaltin,
Sevinc Emre,
Tulin Ibsirlioglu,
Ariela Benigni,
Giuseppe Remuzzi,
Marina Noris
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ABSTRACT: Focal segmental glomerulosclerosis is a kidney disease that is manifested as the nephrotic syndrome. It is often resistant to glucocorticoid therapy and progresses to end-stage renal disease in 50 to 70% of patients. Genetic studies have shown that familial focal segmental glomerulosclerosis is a disease of the podocytes, which are major components of the glomerular filtration barrier. However, the molecular cause in over half the cases of primary focal segmental glomerulosclerosis is unknown, and effective treatments have been elusive.
We performed whole-genome linkage analysis followed by high-throughput sequencing of the positive-linkage area in a family with autosomal recessive focal segmental glomerulosclerosis (index family) and sequenced a newly discovered gene in 52 unrelated patients with focal segmental glomerulosclerosis. Immunohistochemical studies were performed on human kidney-biopsy specimens and cultured podocytes. Expression studies in vitro were performed to characterize the functional consequences of the mutations identified.
We identified two mutations (A159P and Y695X) in MYO1E, which encodes a nonmuscle class I myosin, myosin 1E (Myo1E). The mutations in MYO1E segregated with focal segmental glomerulosclerosis in two independent pedigrees (the index family and Family 2). Patients were homozygous for the mutations and did not have a response to glucocorticoid therapy. Electron microscopy showed thickening and disorganization of the glomerular basement membrane. Normal expression of Myo1E was documented in control human kidney-biopsy specimens in vivo and in glomerular podocytes in vitro. Transfection studies revealed abnormal subcellular localization and function of the A159P-Myo1E mutant. The Y695X mutation causes loss of calmodulin binding and of the tail domains of Myo1E.
MYO1E mutations are associated with childhood-onset, glucocorticoid-resistant focal segmental glomerulosclerosis. Our data provide evidence of a role of Myo1E in podocyte function and the consequent integrity of the glomerular filtration barrier.
New England Journal of Medicine 07/2011; 365(4):295-306. · 53.30 Impact Factor
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ABSTRACT: During the 2009 pandemic the Virology Laboratory of L. Spallanzani, Rome, Italy, adopted a real-time RT-PCR developed by the Centers for Disease Control and Prevention (CDC), Atlanta, Georgia to diagnose pandemic influenza A/H1N1 (H1N1pdm). A new multiplex real-time RT-PCR distributed by Astra Diagnostics, coupled with the extraction system developed and commercialized by Siemens Healthcare Diagnostics (referred to as the RealStar system), was tested for the ability to detect and type influenza A in clinical samples, with particular emphasis on influenza A-positive samples untyped by the CDC method. Seventy-six nasopharyngeal swabs, resulting by the CDC method H1N1pdm (n=7), H3N2 (n=3), and not subtyped (n=66), were re-analysed with the RealStar system. All H3N2 and H1N1pdm-positive samples were correctly identified; among the untyped samples, the RealStar system detected 24/66 (36.4%) H1N1pdm and 1/66 (1.5%) seasonal influenza A. In conclusion, the RealStar system confirmed the results of all the influenza A-positive samples subtyped by the CDC method, and was able to type 37.9% of samples untyped by the CDC method. However, 62.1% of samples, detected as influenza A-positive but not subtyped by the CDC method, were found to be negative by the RealStar system. Further investigation is needed to explain this latter, unexpected, finding.
Journal of virological methods 07/2011; 177(2):193-6. · 2.13 Impact Factor
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ABSTRACT: Human hepatitis E virus (HEV) is considered an emerging pathogen in industrialized countries. The aim of the present study was to contribute to the body of knowledge available on the molecular epidemiology of acute hepatitis E in Italy. Three sets of HEV-specific primers targeting the ORF1 and ORF2 were used to examine serum samples collected from acute hepatitis patients positive for anti-HEV IgG and/or IgM, between 2007 and 2010. Seventeen patients (39.5%) tested HEV RNA-positive: 12 infections, due to genotype 1, were associated with travel to endemic areas (Bangladesh, India and Pakistan), while five infections, due to genotype 3, were presumably autochthonous. Risk factors identified in this group included exposure to raw seafood, pork liver sausages and wild boar. Results from the present study confirm that human HEV infection in Italy is caused by different genotypes, depending on whether the infection is travel-related or autochthonous.
Journal of General Virology 04/2011; 92(Pt 7):1617-26. · 3.36 Impact Factor
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ABSTRACT: To the Editor: The biologic role of amino acid variants at position 222 of the hemagglutinin (HA) gene of pandemic (H1N1) 2009 virus in severe infections has been extensively discussed. A recent series of studies (1-3) confirm the initial suggestions that G or N in this position might confer greater pathogenic potential to the virus than to the wild type. In contrast, their data suggest that no particular pathogenicity is associated with the 222E variant because it occurs at the same frequency in severe and mild infections. Most authors also seem to agree that D222G or N appears sporadically in phylogenetically distant viruses, with limited transmissibility.
Emerging Infectious Diseases 04/2011; 17(4):749-51. · 6.79 Impact Factor
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ABSTRACT: In this work we evaluated plasmacytoid (pDC) and myeloid dendritic (mDC) cells activation before and during anti-HCV treatment in HCV+/HIV+ individuals. HCV+/HIV+ patients received Peg-IFN-α2b subcutaneously for 28 days, followed by oral weight-based ribavirin. DCs activation was evaluated by flow cytometry. Baseline pDC CD80 and CD86 expression was correlated with HIV, but not with HCV viral load. A transient decrease of HIV RNA was found not associated with DC activation. When patients were grouped according to early/sustained virological response (EVR/SVR) to anti-HCV treatment, baseline pDC CD80 and CD86 expression was higher in non-EVR and non-SVR compared to EVR and SVR. Moreover, in responder patients CD80 and CD86 were upregulated by IFN-α. Our data suggest a correlation between DCs activation and response to therapy. These findings could be helpful to better understand the mediators of IFN-α action in HCV+/HIV+ patients and to explore possible exploitation of this knowledge to improve therapeutic response.
Clinical Immunology 02/2011; 138(2):178-86. · 4.05 Impact Factor
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ABSTRACT: Several outbreaks of hepatitis A affecting homosexual men have been reported in Europe. However, the prevalence of HIV infection in patients affected by hepatitis A has not been extensively studied and hepatitis A is not considered as an indicator disease for routine HIV testing.
We retrospectively analyzed all adult cases of acute hepatitis A, reported by the National Institute of Infectious Disease "L. Spallanzani", Rome-Italy, in 2002-2008. Data on HIV infection were obtained by chart review and cross-linkage with laboratory. Information on exposure to risk factors were collected from the standard questionnaire of the Local Health Unit.
We analyzed a total of 473 cases of hepatitis A, 368 (77.2%) males that accounted for 75% of all reported cases in Rome, aged 25-64 years (same gender distribution). During the study period, we diagnosed a high proportion of cases among male individuals (78%). Among the male patients, HIV serology was available for 203/368 (55.2%). The overall HIV prevalence was 15.2% (56/368); it was significantly associated with same gender sex and was significantly higher than that observed among patients with hepatitis B (4.0%).
We found a high HIV prevalence, associated with same gender sex, among adult male patients diagnosed with hepatitis A in the period 2002-2008, except for 2006. Our data suggest that in a low incidence area for hepatitis A, with a constant high proportion of cases among male individuals, all individuals with acute hepatitis A should be routinely offered an HIV test.
Journal of Hepatology 02/2011; 54(6):1102-6. · 9.26 Impact Factor
