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1.
Sci Rep ; 11(1): 9772, 2021 05 07.
Article in English | MEDLINE | ID: mdl-33963235

ABSTRACT

Understanding the SARS-CoV-2 dynamics has been subject of intense research in the last months. In particular, accurate modeling of lockdown effects on human behaviour and epidemic evolution is a key issue in order e.g. to inform health-care decisions on emergency management. In this regard, the compartmental and spatial models so far proposed use parametric descriptions of the contact rate, often assuming a time-invariant effect of the lockdown. In this paper we show that these assumptions may lead to erroneous evaluations on the ongoing pandemic. Thus, we develop a new class of nonparametric compartmental models able to describe how the impact of the lockdown varies in time. Our estimation strategy does not require significant Bayes prior information and exploits regularization theory. Hospitalized data are mapped into an infinite-dimensional space, hence obtaining a function which takes into account also how social distancing measures and people's growing awareness of infection's risk evolves as time progresses. This also permits to reconstruct a continuous-time profile of SARS-CoV-2 reproduction number with a resolution never reached before in the literature. When applied to data collected in Lombardy, the most affected Italian region, our model illustrates how people behaviour changed during the restrictions and its importance to contain the epidemic. Results also indicate that, at the end of the lockdown, around [Formula: see text] of people in Lombardy and [Formula: see text] in Italy was affected by SARS-CoV-2, with the fatality rate being 1.14%. Then, we discuss how the situation evolved after the end of the lockdown showing that the reproduction number dangerously increased in the summer, due to holiday relax, reaching values larger than one on August 1, 2020. Finally, we also document how Italy faced the second wave of infection in the last part of 2020. Since several countries still observe a growing epidemic and others could be subject to other waves, the proposed reproduction number tracking methodology can be of great help to health care authorities to prevent SARS-CoV-2 diffusion or to assess the impact of lockdown restrictions on human behaviour to contain the spread.


Subject(s)
COVID-19/epidemiology , Bayes Theorem , COVID-19/prevention & control , COVID-19/transmission , Communicable Disease Control , Epidemiological Monitoring , Humans , Italy/epidemiology , Models, Statistical , Physical Distancing , SARS-CoV-2/isolation & purification , Seasons , Time Factors
2.
Int J Infect Dis ; 102: 363-368, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33130199

ABSTRACT

BACKGROUND: The high contagiousness and rapid spreading of the coronavirus disease 2019 (COVID-19) has caused a high number of critical to severe life-threatening cases, which required urgent hospital admission and treatment in intensive care units (ICUs). The pandemic has been a tough test for all European national health systems and their capability to provide an adequate reaction. METHODS: The present work aims to reveal correlations between parameters such as COVID-19 incidence, ICU bed occupancy, ICU excess area, and mortality in Italian regions. Public data for the period of March 1 to July 16, 2020, were analyzed using several mathematical and statistical methods. RESULTS: The analysis defined two separate groups of Italian regions. The examined variables considered within these groups were interlinked and dependent on each other. The regions of the two groups shared the same kind of fitted model (linear) explaining mortality as a function of cumulative incidence, but with higher value of the constant in one group, so characterized by a high intrinsic "strength" of the pandemic, certainly playing a major role in the generation of a large number of severe and life-threatening cases. These results are confirmed at European level. Other factors may condition mortality and be linked to incidence, such as ICU saturation and excess. CONCLUSIONS: These quantitative results could be a very helpful tool to set up preventive measures and optimize biomedical interventions before the pandemic, in its recurrent waves, could overcome the reaction capacity of any public health system.


Subject(s)
COVID-19/epidemiology , COVID-19/mortality , Intensive Care Units/statistics & numerical data , COVID-19/therapy , COVID-19/virology , Europe/epidemiology , Hospitalization , Humans , Incidence , Italy/epidemiology , Pandemics , SARS-CoV-2/physiology
3.
Ann Ig ; 32(3): 319-322, 2020.
Article in English | MEDLINE | ID: mdl-32266369
4.
J Hosp Infect ; 102(3): 332-336, 2019 Jul.
Article in English | MEDLINE | ID: mdl-30633945

ABSTRACT

BACKGROUND: Medico-legal conflicts arise when it is difficult to prove the cause of nosocomial infections. AIM: To report an outbreak of patient-to-patient transmission of hepatitis C virus (HCV) through the repeated use of a multi-dose saline flask during the rinsing of central venous catheters. METHODS: Blood samples were taken from each patient for the comparative analysis of their HCV RNA strains. No samples were available for one patient who died before the investigation started. Despite the known lability of HCV RNA, the body was exhumed four months after burial and postmortem samples were collected. HCV RNA was extracted successfully from liver and spleen samples. Genotyping of all the HCV strains was performed by sequence analysis of the 5'NC untranslated region, the E1 core conserved region and the E1/E2 hypervariable region. FINDINGS: Forensic investigators retraced the route used by two ward nurses, when saline catheter flushes were given to 14 patients with each nurse administering to seven patients. The comparative phylogenetic analysis of all case strains identified the deceased patient as the source of contamination to five patients. CONCLUSIONS: This study highlights the value of sequence analysis as a tool for solving medico-legal conflicts. The High Court of Justice found that a health worker's re-use of a contaminated needle resulted in the nosocomial transmission of HCV.


Subject(s)
Cross Infection/epidemiology , Cross Infection/transmission , Disease Transmission, Infectious , Hepacivirus/isolation & purification , Hepatitis C/epidemiology , Hepatitis C/transmission , Adult , Aged , Aged, 80 and over , Cross Infection/mortality , Exhumation , Female , Genotype , Genotyping Techniques , Hepacivirus/classification , Hepacivirus/genetics , Hepatitis C/mortality , Humans , Male , Molecular Epidemiology , RNA, Viral/genetics , RNA, Viral/isolation & purification , Sequence Analysis, DNA
5.
Clin Microbiol Infect ; 25(2): 203-209, 2019 Feb.
Article in English | MEDLINE | ID: mdl-29800674

ABSTRACT

OBJECTIVES: A prospective cohort study was conducted in Italy in order to describe the microbiologic aspects of colonization/infection by carbapenemase-producing Enterobacteriaceae (CPE) in donors and recipients of lung and liver transplants and the possible CPE transmission from donors to recipients. METHODS: Between 15 January 2014 and 14 January 2015, all recipients of solid organ transplants (SOT) at ten lung and eight liver transplantation centres and the corresponding donors were enrolled. Screening cultures to detect CPE were performed in donors, and screening and clinical cultures in recipients with a 28-day microbiologic follow-up after receipt of SOT. Detection of carbapenemase genes by PCR, genotyping by multilocus sequence typing, and pulsed-field gel electrophoresis and whole-genome sequencing were performed. RESULTS: Of 588 screened donors, 3.4% were colonized with CPE. Of the liver first transplant recipients (n = 521), 2.5% were colonized before receipt of SOT and 5% acquired CPE during follow-up. CPE colonization was higher in lung first transplant recipients (n = 111, 2.7% before SOT and 14.4% after SOT). CPE infections occurred in 1.9% and 5.3% of liver or lung recipients, respectively. CPE isolates were mostly Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae belonging to CG258. Three events of donor-recipient CPE transmission, confirmed by whole-genome sequencing and/or pulsed-field gel electrophoresis, occurred in lung recipients: two involving K. pneumoniae sequence type 512 and one Verona integron-encoded metallo-ß-lactamase (VIM)-producing Enterobacter aerogenes. CONCLUSIONS: This study showed a low risk of donor-recipient CPE transmission, indicating that donor CPE colonization does not necessarily represent a contraindication for donation unless colonization regards the organ to be transplanted. Donor and recipient screening remains essential to prevent CPE transmission and cross-infection in transplantation centres.


Subject(s)
Bacterial Proteins/metabolism , Carbapenem-Resistant Enterobacteriaceae , Enterobacteriaceae Infections/microbiology , Liver Transplantation/adverse effects , Lung Transplantation/adverse effects , beta-Lactamases/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Cohort Studies , Drug Resistance, Bacterial , Enterobacteriaceae Infections/epidemiology , Female , Humans , Infant , Italy/epidemiology , Male , Middle Aged , Prospective Studies , Tissue Donors , Transplant Recipients , Young Adult
6.
Epidemiol Infect ; 145(16): 3375-3384, 2017 12.
Article in English | MEDLINE | ID: mdl-29145911

ABSTRACT

We determined the hepatitis E virus (HEV) seroprevalence and detection rate in commercial swine herds in Italy's utmost pig-rich area, and assessed HEV seropositivity risk in humans as a function of occupational exposure to pigs, diet, foreign travel, medical history and hunting activities. During 2011-2014, 2700 sera from 300 swine herds were tested for anti-HEV IgG. HEV RNA was searched in 959 faecal pools from HEV-seropositive herds and in liver/bile/muscle samples from 179 pigs from HEV-positive herds. A cohort study of HEV seropositivity in swine workers (n = 149) was also performed using two comparison groups of people unexposed to swine: omnivores (n = 121) and vegetarians/vegans (n = 115). Herd-level seroprevalence was 75·6% and was highest in farrow-to-feeder herds (81·6%). Twenty-six out of 105 (24·8%) herds had HEV-positive faecal samples (25 HEV-3, one HEV-4). Only one bile sample tested positive. HEV seropositivity was 12·3% in swine workers, 0·9% in omnivores and 3·0% in vegetarians/vegans. Factors significantly associated with HEV seropositivity were occupational exposure to pigs, travel to Africa and increased swine workers' age. We concluded that HEV is widespread in Italian swine herds and HEV-4 circulation is alarming given its pathogenicity, with those occupationally exposed to pigs being at increased risk of HEV seropositivity.


Subject(s)
Farmers/statistics & numerical data , Hepatitis E virus/immunology , Hepatitis E/epidemiology , Hepatitis E/virology , Occupational Exposure , Adult , Aged , Animals , Bile , Feces , Female , Hepatitis Antibodies/blood , Hepatitis E/immunology , Hepatitis E/veterinary , Humans , Immunoglobulin G/blood , Italy/epidemiology , Male , Middle Aged , Muscles , RNA, Viral/blood , Risk Factors , Seroepidemiologic Studies , Swine , Young Adult
7.
Eur J Clin Microbiol Infect Dis ; 35(8): 1247-58, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27234593

ABSTRACT

Laboratory-acquired infections due to a variety of bacteria, viruses, parasites, and fungi have been described over the last century, and laboratory workers are at risk of exposure to these infectious agents. However, reporting laboratory-associated infections has been largely voluntary, and there is no way to determine the real number of people involved or to know the precise risks for workers. In this study, an international survey based on volunteering was conducted in biosafety level 3 and 4 laboratories to determine the number of laboratory-acquired infections and the possible underlying causes of these contaminations. The analysis of the survey reveals that laboratory-acquired infections have been infrequent and even rare in recent years, and human errors represent a very high percentage of the cases. Today, most risks from biological hazards can be reduced through the use of appropriate procedures and techniques, containment devices and facilities, and the training of personnel.


Subject(s)
Biomedical Research , Laboratories , Occupational Diseases , Occupational Exposure , Biomedical Research/standards , Biomedical Research/statistics & numerical data , Containment of Biohazards , Cross-Sectional Studies , Humans , Laboratories/standards , Laboratories/statistics & numerical data , Occupational Diseases/epidemiology , Occupational Diseases/microbiology , Occupational Diseases/prevention & control , Occupational Diseases/virology , Occupational Exposure/prevention & control , Occupational Exposure/standards , Occupational Exposure/statistics & numerical data , Personal Protective Equipment/standards , Personal Protective Equipment/statistics & numerical data , Risk Assessment , Safety , Surveys and Questionnaires
8.
Ann Ig ; 28(1): 15-24, 2016.
Article in English | MEDLINE | ID: mdl-26980506

ABSTRACT

BACKGROUND: From 2007, in the Veneto Region (Italy), a surveillance system for invasive pneumococcal diseases (IPD) was implemented to estimate the regional epidemiology of IPD and to evaluate the impact of 13-valent pneumococcal conjugate vaccine (PCV13) vaccination. METHODS: Data were collected from 2007 to 2014 and the total, annual and age-specific IPD notification rates were calculated. A Poisson regression model was used to identify the possible risk factors for developing IPD. RESULTS: A total of 713 IPD cases were notified and the overall IPD notification rate was equal to 2.0 cases per 100,000 population (95% CI: 1.7-2.1), with an increasing trend between 2007 and 2014. The pneumococcal serotypes were identified in 608 (85.3%) isolates from biological specimens, and the most distributed serotypes were those contained in PCV13. Children <5 year-old and the adults over 65 year-old showed the highest PCV13 vaccine-type IPD notification rate, equal to 2.7/100,000 and 2.8/100,000, respectively. The risk to develop IPD was greater in children aged <5 years (RR = 8.9, 95% CI: 5.1-15.9; p<0.0001) and in adults aged >65 years (RR = 4.3, 95% CI: 2.7-6.9; p<0.0001), especially in males > 65 years of age (RR = 1.7, 95% CI: 1.0-2.8; p = 0.042). The invasive pneumococcal disease was mainly caused by the PCV13 serotypes (RR = 2.9, 95%CI: 2.3-3.9; p<0.0001), principally after the PCV13 introduction (RR = 2.3, 95% CI: 1.4-3.8; p<0.001). In spite of that, a significant reduction of the overall IPD incidence is evident in the period following the PCV13 vaccine introduction (RR = 0.4, 95% CI: 0.3-0.5; p<0.0001), particularly in children aged <5 years (RR = 0.3, 95% CI: 0.2-0.7; p = 0.002), demonstrating the real efficacy of PCV13 immunization for children. CONCLUSIONS: In the Veneto Region, the surveillance system has allowed to describe the detailed epidemiological profile of invasive pneumococcal disease, pointing out that the most circulating pneumococcal serotypes were those contained in the PCV13 vaccine.


Subject(s)
Pneumococcal Infections/epidemiology , Pneumococcal Infections/prevention & control , Pneumococcal Vaccines/administration & dosage , Vaccination , Adolescent , Adult , Age Distribution , Aged , Child , Child, Preschool , Female , Humans , Incidence , Italy/epidemiology , Male , Middle Aged , Pneumococcal Infections/immunology , Pneumococcal Vaccines/immunology , Population Surveillance , Prospective Studies , Risk Factors , Sex Distribution , Vaccination/methods
9.
Clin Microbiol Infect ; 21(12): 1122.e1-10, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26235197

ABSTRACT

In recent years, West Nile virus (WNV) lineage 2 has been spreading and causing disease outbreaks in humans and animals in Europe. In order to characterize viral diversity, we performed full-length genome sequencing of WNV lineage 2 from human samples collected during outbreaks in Italy and Greece in 2013 and 2014. Phylogenetic analysis showed that these WNV lineage 2 genomes belonged to a monophyletic clade derived from a single introduction into Europe of the prototype Hungarian strain. Correlation of phylogenetic data with geospatial information showed geographical clustering of WNV genome sequences both in Italy and in Greece, indicating that the virus had evolved and diverged during its dispersal in Europe, leading to the emergence of novel genotypes, as it adapted to local ecological niches. These genotypes carried divergent conserved amino acid substitutions, which might have been relevant for viral adaptation, as suggested by selection pressure analysis and in silico and experimental modelling of sequence changes. In conclusion, the results of this study provide further information on WNV lineage 2 transmission dynamics in Europe, and emphasize the need for WNV surveillance activities to monitor viral evolution and diversity.


Subject(s)
Disease Outbreaks , RNA, Viral/genetics , West Nile Fever/epidemiology , West Nile virus/classification , West Nile virus/genetics , Amino Acid Substitution , Evolution, Molecular , Genome, Viral , Greece , Humans , Italy , Models, Molecular , Phylogeny , Phylogeography , Sequence Analysis, RNA , West Nile Fever/transmission
10.
Clin Microbiol Infect ; 21(3): 288.e5-7, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25658542

ABSTRACT

In a longitudinal study on 181 naïve patients who responded to therapy (mean follow-up 4 years), high baseline human immunodeficiency virus (HIV)-RNA values correlated with high levels of cellular HIV-DNA at all time points (p < 0.0001, p 0.045, p 0.0055, and p 0.0025, respectively) and negatively correlated with undetectable residual viremia (URV; <2.5 copies/mL) at T1, T2, and T3 (p 0.026, p 0.0149, and p 0.0002, respectively). Baseline high HIV-DNA levels predicted the persistence of high values (p 0.0001) and negatively correlated with URV (p 0.0254, p 0.0481, and p 0.0085). These results suggest that baseline viral load, cellular HIV-DNA, and URV were strongly correlated over long-term follow-up of antiretroviral therapy responders.


Subject(s)
Antiretroviral Therapy, Highly Active , HIV Infections/drug therapy , HIV Infections/virology , HIV-1/genetics , Leukocytes, Mononuclear/virology , Viral Load , Viremia , Adult , CD4 Lymphocyte Count , DNA, Viral , Female , Follow-Up Studies , Genotype , HIV Infections/epidemiology , Humans , Longitudinal Studies , Male , Middle Aged , Treatment Outcome
11.
J Cell Physiol ; 230(8): 1794-806, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25510868

ABSTRACT

Similar to phosphorylation, transient conjugation of ubiquitin to target proteins (ubiquitination) mediated by the concerted action of ubiquitin ligases and de-ubiquitinating enzymes (DUBs) can affect substrate function. As obligate intracellular parasites, viruses rely on different cellular pathways for their own replication and the well conserved ubiquitin conjugating/de-conjugating system is not an exception. Viruses not only usurp the host proteins involved in the ubiquitination/de-ubiquitination process, but they also encode their own ubiquitin ligases and DUBs. Here we report that an N-terminal variant of the herpes simplex virus (HSV) type-1 large tegument protein VP1/2 (VP1/2(1-767)), encompassing an active DUB domain (herpesvirus tegument ubiquitin specific protease, htUSP), and TSG101, a component of the endosomal sorting complex required for transport (ESCRT)-I, functionally interact. In particular, VP1/2(1-767) modulates TSG101 ubiquitination and influences its intracellular distribution. Given the role played by the ESCRT machinery in crucial steps of both cellular pathways and viral life cycle, the identification of TSG101 as a cellular target for the HSV-1 specific de-ubiquitinating enzyme contributes to the clarification of the still under debate function of viral encoded DUBs highly conserved throughout the Herpesviridae family.


Subject(s)
DNA-Binding Proteins/metabolism , Endosomal Sorting Complexes Required for Transport/metabolism , Host-Parasite Interactions/physiology , Simplexvirus/pathogenicity , Transcription Factors/metabolism , Ubiquitin-Specific Proteases/metabolism , Viral Proteins/metabolism , Animals , Chlorocebus aethiops , Humans , Immunoprecipitation , Microscopy, Confocal , Real-Time Polymerase Chain Reaction , Simplexvirus/metabolism , Ubiquitination , Vero Cells
12.
Gene Ther ; 21(3): 272-81, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24430237

ABSTRACT

Corneal graft rejection is a major problem in chronic herpetic keratitis (HK) patients with latent infection. A new class of antiviral agents targeting latent and active forms of herpes simplex virus type 1 (HSV-1) is importantly required. Meganucleases are sequence-specific homing endonucleases capable of inducing DNA double-strand breaks. A proof-of-concept experiment has shown that tailor-made meganucleases are efficient against HSV-1 in vitro. To take this work a step forward, we hypothesized that the pre-treatment of human corneas in eye banks using meganuclease-encoding vectors will allow HK patients to receive a medicated cornea to resist the recurrence of the infection and the common graft rejection problem. However, this strategy requires efficient gene delivery to human corneal endothelium. Using recombinant adeno-associated virus, serotype 2/1 (rAAV2/1), efficient gene delivery of a reporter gene was demonstrated in human corneas ex vivo. The optimum viral dose was 3.7 × 10(11) VG with an exposure time of 1 day, followed by 6 days incubation in de-swelling medium. In addition, 12 days incubation can result in transgene expression in excess of 70%. Using similar transduction conditions, meganuclease transgene expression was detected in 39.4% of the endothelial cells after 2 weeks in culture. Reduction of the total viral load in the media and the endothelial cells of corneas infected with HSV-1 was shown. Collectively, this work provides information about the optimum conditions to deliver genetic material to the cornea, and demonstrates for the first time the expression of meganuclease in human corneas ex vivo and its antiviral activity. In conclusion, we demonstrate that the treatment of human corneas in eye banks before transplantation is a new approach to address the unmet clinical needs in corneal diseases.


Subject(s)
Cornea/metabolism , Deoxyribonuclease I/genetics , Viral Proteins/genetics , Deoxyribonuclease I/metabolism , Dependovirus/genetics , Dependovirus/metabolism , Gene Transfer Techniques , Genes, Reporter/genetics , Herpesvirus 1, Human/enzymology , Humans , In Vitro Techniques , Viral Proteins/metabolism
13.
Int J Hyg Environ Health ; 217(1): 17-22, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23540488

ABSTRACT

Influenza virus spreads via small particle aerosols, droplets and fomites, and since it can survive for a short time on surfaces, can be introduced into the nasal mucosa before it loses infectivity. The hypothiocyanite ion (OSCN-), product of the lactoperoxidase/H2O2/SCN- system of central airways, is emerging as an important molecule for innate defense mechanism against bacteria, fungi and viruses. Here we demonstrated that OSCN(-) displays virucidal activity in vitro against the A/H1N1 2009 pandemic influenza virus. The concentration required to inhibit viral replication by 50% was 2 µM when virus were challenged directly with OSCN- before cell inoculation. These values were even lower when inoculated cells were maintained in contact with enzyme free-OSCN- in the culture medium. The last experimental conditions better reflect those of tracheobronchial mucosa, where HOSCN/OSCN- is retained in the air-liquid interface and inactivates both the viruses approaching the epithelium from outside and those released from the inoculated cells after the replication cycle. Importantly no OSCN- cytotoxicity was observed in the cellular system employed. The lack of toxicity in humans and the absence of damage on surfaces of fomites suggest a potential use of OSCN- to avoid mucosal and environmental transmission of influenza virus. Since hypothiocyanite is normally present in human airways a low risk of viral resistance is envisaged. In vivo confirmatory studies are needed to evaluate the appropriate dose, regimen and formulation.


Subject(s)
Antiviral Agents/pharmacology , Influenza A Virus, H1N1 Subtype/drug effects , Thiocyanates/pharmacology , Animals , Cell Line , Dogs , Influenza A Virus, H1N1 Subtype/physiology , Pandemics , Virus Internalization/drug effects
14.
Euro Surveill ; 18(38)2013 Sep 19.
Article in English | MEDLINE | ID: mdl-24084339

ABSTRACT

A human outbreak of West Nile virus (WNV) infection caused by WNV lineage 2 is ongoing in northern Italy. Analysis of six WNV genome sequences obtained from clinical specimens demonstrated similarities with strains circulating in central Europe and Greece and the presence of unique amino acid changes that identify a new viral strain. In addition, WNV lineage 1 Livenza, responsible for a large outbreak in north-eastern Italy in 2012, was fully sequenced from a blood donor during this 2013 outbreak.


Subject(s)
RNA, Viral/genetics , West Nile Fever/genetics , West Nile virus/classification , West Nile virus/genetics , Base Sequence , Disease Outbreaks , Genome , Humans , Italy/epidemiology , Molecular Epidemiology , Phylogeny , West Nile Fever/diagnosis , West Nile Fever/epidemiology , West Nile Fever/virology
15.
Clin Microbiol Infect ; 19(10): E428-34, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23573945

ABSTRACT

Accurate HPV typing is essential for evaluation and monitoring of HPV vaccines, for second-line testing in cervical cancer screening, and in epidemiological surveys. In this study, we set up and assessed in clinical samples a new HPV typing method based on 454 next-generation sequencing (NGS) of HPV L1 amplicons, generated by using a modified PGMY primer set with improved sensitivity for some HPV types that are not targeted by standard PGMY primers. By using a median 12 800-fold coverage, the NGS method allowed us to correctly identify all high-risk HPV types, in either single or multiple infections, with a sensitivity of 50 genome equivalents, as demonstrated by testing WHO LabNet EQA sample panels. Analysis of mixtures of HPV16- and HPV18-positive cell lines demonstrated that the NGS method could reproducibly quantify the proportion of each HPV type in multiple infections in a wide dynamic range. Testing of HPV-positive clinical samples showed that NGS could correctly identify a high number of HPV types in multiple infections. The NGS method was also effective in the analysis of a set of cervical specimens with discordant results at hybrid capture 2 and line probe assays. In conclusion, a new HPV typing method based on 454 pyrosequencing was set up. This method was sensitive, specific, quantitative and precise in both single and multiple infections. It could identify a wide range of HPV types and might potentially discover new HPV types.


Subject(s)
High-Throughput Nucleotide Sequencing/methods , Molecular Typing/methods , Papillomaviridae/classification , Papillomaviridae/genetics , Papillomavirus Infections/virology , Genitalia/virology , Genotype , Humans , Papillomaviridae/isolation & purification , Papillomavirus Infections/diagnosis , Reproducibility of Results , Sensitivity and Specificity
16.
Euro Surveill ; 17(36): 20260, 2012 Sep 06.
Article in English | MEDLINE | ID: mdl-22971328

ABSTRACT

In July-September 2012, one month earlier than in previous years, 13 confirmed human cases of West Nile virus infection were diagnosed in northern Italy, including five with neuroinvasive disease, three with West Nile fever, and five West Nile virus (WNV)-positive blood donors. In nine cases, the presence of the WNV lineage 1a Livenza strain, characterised in 2011, was ascertained. Symptomatic patients had prolonged viruria with high viral load.


Subject(s)
Disease Outbreaks , RNA, Viral/genetics , West Nile Fever/virology , West Nile virus/genetics , Blood Donors , Follow-Up Studies , Humans , Italy/epidemiology , Population Surveillance/methods , Real-Time Polymerase Chain Reaction , Sequence Analysis , Viral Load , West Nile Fever/epidemiology , West Nile Fever/genetics , West Nile virus/isolation & purification
17.
Clin Microbiol Infect ; 18(12): E541-4, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23004685

ABSTRACT

During 2008-2009, several human cases of WNV disease caused by an endemic lineage 1a strain were reported in areas surrounding the Po river in north-eastern Italy. Since 2010, cases have been recorded in nearby northern areas, where, in 2011, both lineage 1a and 2 were detected. We describe here two new WNV complete genome sequences from human cases of WNV infection occurring in 2011 in the Veneto Region. Phylogenetic analysis showed that both genome sequences belonged to lineage 1a and were related to WNV strains of the Western Mediterranean subtype. The novel WNV genomes had high nucleotide and amino acid sequence divergence from each other and from the WNV strain circulating in Italy in 2008-2009. The presence of different WNV strains in a relatively small geographical area is a novel finding with unpredictable impact on human disease that requires further investigation.


Subject(s)
Genome, Viral , RNA, Viral/genetics , Sequence Analysis, DNA , West Nile Fever/virology , West Nile virus/genetics , Genetic Variation , Genotype , Humans , Italy , Molecular Sequence Data , Phylogeny , West Nile virus/classification , West Nile virus/isolation & purification
18.
Euro Surveill ; 17(31)2012 Aug 02.
Article in English | MEDLINE | ID: mdl-22874456

ABSTRACT

We report here the first blood donation positive for West Nile virus (WNV) by nucleic acid amplification testing collected in north-eastern Italy in July 2012.Partial sequencing of the WNV RNA demonstrated identity with a WNV lineage 1a genome identified in the same area in 2011 and divergence from the strain responsible for the outbreak in northern Italy in 2008­09. These data indicate that WNV activity in northern Italy is occurring earlier than expected and that different WNV strains are circulating.


Subject(s)
Blood Donors , RNA, Viral/genetics , West Nile Fever/virology , West Nile virus/genetics , Endemic Diseases , Humans , Italy/epidemiology , Nucleic Acid Amplification Techniques , Phylogeny , Population Surveillance , Sequence Analysis , West Nile Fever/epidemiology , West Nile Fever/genetics
19.
J Neuroimmune Pharmacol ; 7(3): 665-72, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22585413

ABSTRACT

The risk of progressive multifocal leukoencephalopathy (PML) in patients treated with natalizumab for multiple sclerosis (MS) is a serious concern. The presence of anti-JC virus antibodies is a risk factor for PML development, but 2.5 % of the patients result falsely-negative, while the prognostic relevance of testing JCV-DNA in biological fluids of treated patients is debated. Aim of this work was to evaluate the utility of testing JCV-DNA, together with anti-JCV antibodies, in biological samples of treated patients as a tool for PML risk stratification. 126 subjects from 5 MS Centers in Italy were included in the study. We performed a cross-sectional study in 63 patients testing JCV-DNA in blood, peripheral blood cells and urine. We longitudinally assessed the presence of JCV-DNA in a cohort of 33 subjects, one of which developed PML. We could test retrospectively serum samples from another PML case occurred during natalizumab therapy. Anti-JCV antibodies and urinary JCV-DNA were both tested in 73 patients. No changes in JCV-DNA status occurred during natalizumab treatment. The subject who developed PML in the longitudinal cohort had detectable JCV-DNA in urine at all time-points while serum or blood from both PML patients were always negative before the onset of disease and, in one case, after. Four subjects with JCV-DNA in urine and undetectable anti-JCV antibodies were retested for anti-JCV antibodies and three out of four resulted positive. In conclusion, testing JCV-DNA in urine is complementary to testing anti-JCV antibodies in identifying patients at risk of PML.


Subject(s)
Antibodies, Monoclonal, Humanized/therapeutic use , DNA, Viral/urine , JC Virus/metabolism , Leukoencephalopathy, Progressive Multifocal/diagnosis , Leukoencephalopathy, Progressive Multifocal/urine , Adult , Biomarkers/urine , Cross-Sectional Studies , Diagnostic Tests, Routine , Female , Humans , Leukoencephalopathy, Progressive Multifocal/drug therapy , Longitudinal Studies , Male , Middle Aged , Natalizumab , Retrospective Studies , Risk Factors , Young Adult
20.
Res Vet Sci ; 93(2): 872-8, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22083104

ABSTRACT

Between 2001 and 2010, 244 clinically suspected cases of bovine spongiform encephalopathy (BSE) were reported in Italy. This report summarizes the neuropathological findings in cattle displaying clinical signs consistent with a diagnosis of BSE. All animal specimens were submitted for confirmatory testing; samples testing negative underwent neuropathological examination to establish the differential diagnosis. Immunohistochemistry for scrapie prion protein (PrPSc) at the level of frontal cortex was carried out to exclude atypical BSE. Neuropathological changes were detected in 34.9% of cases; no histological lesions were found in 52.3% of subjects; 12.8% of samples were found unsuitable for analysis. BSE was detected in one case, but no cases of atypical BSE were observed. This study identified the diseases most commonly encountered in the differential diagnosis of BSE; furthermore, it demonstrated that the surveillance system is necessary for monitoring neuropathological disease in cattle and for the detection of BSE cases.


Subject(s)
Encephalopathy, Bovine Spongiform/epidemiology , Animals , Brain/pathology , Cattle , Encephalopathy, Bovine Spongiform/pathology , Female , Italy/epidemiology , Male , Population Surveillance , Real-Time Polymerase Chain Reaction/veterinary , Reverse Transcriptase Polymerase Chain Reaction/veterinary
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