40 research outputs found
HCV full-length genome reconstruction with sequence independent amplification combined with next generation sequencing
A case of dengue type 3 virus infection imported from Africa to Italy, October 2009.
In October 2009, a traveller returning from Africa to Italy was hospitalised with symptoms suggestive of a haemorrhagic fever of unknown origin. The patient was immediately placed in a special biocontainment unit until laboratory investigations confirmed the infection to be caused by a dengue serotype 3 virus. This case reasserts the importance of returning travellers as sentinels of unknown outbreaks occurring in other countries, and highlights how the initial symptoms of dengue fever resemble those of other haemorrhagic fevers, hence the importance of prompt isolation of patients until a final diagnosis is reached
Characterization of the patterns of drug-resistance mutations in newly diagnosed HIV-1 infected patients naĂŻve to the antiretroviral drugs
<p>Abstract</p> <p>Background</p> <p>The transmission of HIV-1 drug-resistant strains in drug naive patients may seriously compromise the efficacy of a first-line antiretroviral treatment. To better define this problem, a study in a cohort of newly diagnosed HIV-1 infected individuals has been conducted. This study is aimed to assess the prevalence and the patterns of the mutations recently associated with transmitted drug resistance in the reverse transcriptase (RT) and in protease (PR) of HIV-1.</p> <p>Methods</p> <p>Prevalence of transmitted drug resistant strains is determined in 255 newly diagnosed HIV-1 infected patients enrolled in different counselling and testing (CT) centres in Central Italy; the Avidity Index (AI) on the first available serum sample is also used to estimate time since infection. Logistic regression models are used to determine factors associated with infection by drug resistant HIV-1 strains.</p> <p>Results</p> <p>The prevalence of HIV-1 strains with at least one major drug resistance mutation is 5.9% (15/255); moreover, 3.9% (10/255) of patients is infected with HIV nucleoside reverse transcriptase inhibitor (NRTI)-resistant viruses, 3.5% (9/255) with HIV non-NRTI-resistant viruses and 0.4% (1/255) with HIV protease inhibitor (PI)-resistant viruses. Most importantly, almost half (60.0%) of patients carries HIV-1 resistant strains with more than one major drug resistance mutation. In addition, patients who had acquired HIV through homosexual intercourses are more likely to harbour a virus with at least one primary resistance mutation (OR 7.7; 95% CI: 1.7–35.0, P = 0.008).</p> <p>Conclusion</p> <p>The prevalence of drug resistant HIV-1 strains among newly diagnosed individuals in Central Italy is consistent with the data from other European countries. Nevertheless, the presence of drug-resistance HIV-1 mutations in complex patterns highlights an additional potential risk for public health and strongly supports the extension of wide genotyping to newly diagnosed HIV-1 infected patients.</p
Clinical Features, Cardiovascular Risk Profile, and Therapeutic Trajectories of Patients with Type 2 Diabetes Candidate for Oral Semaglutide Therapy in the Italian Specialist Care
Introduction: This study aimed to address therapeutic inertia in the management of type 2 diabetes (T2D) by investigating the potential of early treatment with oral semaglutide. Methods: A cross-sectional survey was conducted between October 2021 and April 2022 among specialists treating individuals with T2D. A scientific committee designed a data collection form covering demographics, cardiovascular risk, glucose control metrics, ongoing therapies, and physician judgments on treatment appropriateness. Participants completed anonymous patient questionnaires reflecting routine clinical encounters. The preferred therapeutic regimen for each patient was also identified. Results: The analysis was conducted on 4449 patients initiating oral semaglutide. The population had a relatively short disease duration (42%  60% of patients, and more often than sitagliptin or empagliflozin. Conclusion: The study supports the potential of early implementation of oral semaglutide as a strategy to overcome therapeutic inertia and enhance T2D management
Metagenomic approach for discovering new pathogens in infection disease outbreaks
Viruses represent the most abundant biological components on earth.They can be found in every environment, from deep layers of oceans to animal bodies.Although several viruses have been isolated and sequenced, in each environment there are millions of different types of viruses that have not been identified yet.The advent of nextgeneration sequencing technologies with their high throughput capabilities make possible to study in a single experiment all the community of microorganisms present in a particular sample “microbioma”.They made more feasible the application of the metagenomic approach, by which it is also possible to discover and identify new pathogens, that may pose a threat to public health.This paper summarizes the most recent applications of nextgeneration sequencing to discover new viral pathogens during the occurrence of infection disease outbreaks
Comparison of two NGS platforms for metagenomic analysis of clinical samples
The advent of next generation sequencing platforms has greatly improved the direct assessment of human metagenomes permitting to obtain results in a time-frame compatible with diagnostic needs. The performances of two platforms, namely Roche 454 GS FLX+ and Illumina MiSeq, have been compared using a metagenomic approach aimed at the identification of the pathogenic agent, an RNA virus in this case, in a clinical sample. Both platforms resulted able to correctly identify the H1N1 virus in the sample, but also provided a similar overview of the microbial community. The detailed analysis of two very different but clinically relevant microorganisms that often co-infect patients, H1N1 and Streptococcus pneumoniae, showed differences in terms of depth of coverage and genome coverage, and showed that some genomic regions are more frequently represented in Illumina sequencing reads, while others in the 454 reads. These findings indicate that the platforms and the corresponding analysis procedures permit the high quality assembly of the H1N1 virus and the identification of the complex microbial community in the sample, supporting the usage of these approaches in a clincal setting
Whole Genome Characterization of Orthopoxvirus (OPV) Abatino, a Zoonotic Virus Representing a Putative Novel Clade of Old World Orthopoxviruses
Orthopoxviruses (OPVs) are diffused over the complete Eurasian continent, but previously described strains are mostly from northern Europe, and few infections have been reported from Italy. Here we present the extended genomic characterization of OPV Abatino, a novel OPV isolated in Italy from an infected Tonkean macaque, with zoonotic potential. Phylogenetic analysis based on 102 conserved OPV genes (core gene set) showed that OPV Abatino is most closely related to the Ectromelia virus species (ECTV), although placed on a separate branch of the phylogenetic tree, bringing substantial support to the hypothesis that this strain may be part of a novel OPV clade. Extending the analysis to the entire set of genes (coding sequences, CDS) further substantiated this hypothesis. In fact the genome of OPV Abatino included more CDS than ECTV; most of the extra genes (mainly located in the terminal genome regions), showed the highest similarity with cowpox virus (CPXV); however vaccinia virus (VACV) and monkeypox virus (MPXV) were the closest OPV for certain CDS. These findings suggest that OPV Abatino could be the result of complex evolutionary events, diverging from any other previously described OPV, and may indicate that previously reported cases in Italy could represent the tip of the iceberg yet to be explored.Peer Reviewe
Critical reappraisal of the A226V mutation in Chikungunya outbreaks: possible role in increased pathogenesis?
CHIKV is a mosquito-transmitted alphavirus responsible for the first autochthonous Italian outbreak in 2007.We previously analyzed 7 CHIKV isolates (5 imported and 2 autochthonous) with respect to the presence of A226V mutation in E1gp. All the isolates showed this mutation except the one imported from India in 2006. Since this mutation has been associated with enhanced replication and fitness in A. albopictus vector, we investigated the possible involvement of A226V mutation in enhanced infection capability in primate cells. To this aim,Vero E6 and C6/36 cells were infected with two CHIKV isolates, one carrying the A226V mutation and one wild type, using single replication cycle conditions. Progeny virus was measured by both quantitative real time RT-PCR and viral infectivity assay. No significant differences were observed between the two isolates either in terms of replication kinetic or in virus yield, on both Vero E6 and C6/36 cells. Moreover, experiment of inhibition of virus replication were performed for both isolates on Vero E6 cells using increasing amounts of recombinant IFN-alpha and virus yield was measured. A dose-dependent inhibition of virus yield for both CHIK isolates was observed, with a different sensitivity to IFN-alpha between the isolate carrying the A226V mutation and the wild type one. Our results suggest i) that A226V mutation does not influence replication ability in both host species, when using single replication cycle conditions; ii) the differences between wild type and mutated strains may be due to different sensitivity and/or activation ability of innate immune mechanisms