Dolutegravir-based regimen maintains virological success in a patient with archived mutations to integrase inhibitors

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Seroprevalence of group B Coxsackieviruses: retrospective study in an Italian population

Purpose Group B Coxsackieviruses (CVB) include 6 serotypes (B1‐6) responsible for a wide range of clinical diseases. Since no recent seroepidemiologic data are available in Italy, the study aim was to investigate CVB seroprevalence in a wide Italian population. Methods The study retrospectively included 2,459 subjects referring to a large academic hospital in Rome (Italy) in the period 2004‐2016. Seroprevalence rates and neutralizing antibodies (nAb) titers were evaluated in relation to years of observation and subjects’ characteristics. Results Positivity for at least one serotype was detected in 69.1% of individuals. Overall, the prevalent serotype was B4, followed by B3 (33.3%), B5 (26.2%), B1 (12.7%), B2 (11.0%), and B6 (1.7%). For B2, a significant decrease in seroprevalence over years was observed. Positivity to at least one virus was 25.2% in children aged 0‐2 years, but significantly increased in pre‐school (3‐5 yr) (50.3%) and school (6‐10 yr) children (70.4%). Higher nAb responses for B3 and B4 were observed in children aged 3‐5 years. Conclusion A high overall CVB prevalence was found. Type‐specific variations in prevalence over time probably reflect the fluctuations in circulation typical of Enteroviruses. Children are at greater risk for CVB infection given the high number of seronegative subjects aged 0‐10 years

Humoral and T-cell mediated response after administration of mRNA vaccine BNT162b2 in frail populations

Patients with frailty are considered to be at greater risk to get severe infection from SARS-CoV-2. One of the most effective strategies is vaccination. In our study we evaluated both the humoral immune response elicited by the vaccination at different time points, and the T-cell response in terms of interferon (IFN)-γ production in frail patients and healthy donors. Fifty-seven patients (31 patients undergoing hemodialysis and 26 HIV positive subjects) and 39 healthcare workers were enrolled. All participants received two doses of the mRNA vaccine BNT162b2. Healthcare workers showed a significantly higher antibody titer than patients twenty-one days after the first dose (p < 0.001). From the same time point we observed for both groups a decay of the antibody levels with a steeper slope of decline in the patients group. Regarding T-cell response the only significant difference between non-reactive and reactive subjects was found in median antibody levels, higher in the responders group than in non-responders. The healthcare workers seem to better respond to the vaccination in terms of antibodies production; the lack of T-cell response in about 50% of the participants seems to suggest that in our study population both humoral and cell-mediated response decline over time remarking the importance of the booster doses, particularly for frail patients

transmitted drug resistance mutations and trends of hiv 1 subtypes in treatment naive patients a single centre experience

Abstract Background Transmitted drug resistances (TDRs) and HIV-1 diversity could affect treatment efficacy and clinical outcomes. Here we describe the circulating viral subtypes and estimate the prevalence of resistance among drug naive patients attending Sapienza University Hospital in Rome from 2006-2017. Methods Genotypic resistance test (GRT) was performed on 668 ART-naive patients. GRT were conducted in integrase (n = 52), protease and reverse transcriptase (n = 668) sequences. Results Twenty-one different subtypes and Circulating Recombinant Forms (CRFs) were identified. Subtype B was the most common (67%), followed by CRF02_AG (8.3%), subtypes C and F (6%). We found a significantly increased overtime in the proportion of non-B strains and in the rates of non-Italian patients (p  Minor or accessory INSTI mutations were detected in 17.3% of patients. No significant decrease of TDR prevalence was documented overtime. Conclusion The significant increase of non-B subtypes suggests that the molecular epidemiology of HIV-1 is changing. The detection of a major INSTI mutation in two naive patients highlights the importance of performing GRT before commencing treatment. This finding and the lack of a significant reduction of TDR underline the importance of a continuous surveillance of resistance mutations

The Many Faces of Immune Activation in HIV-1 Infection: A Multifactorial Interconnection

Chronic immune activation has a significant role in HIV-1 disease pathogenesis and CD4+ T-cell depletion. The causes of chronic inflammation and immune activation are incompletely understood, but they are likely multifactorial in nature, involving both direct and indirect stimuli. Possible explanations include microbial translocation, coinfection, and continued presence of competent replicating virus. In fact, long-term viral suppression treatments are unable to normalize elevated markers of systemic immune activation. Furthermore, high levels of pro-inflammatory cytokines increase susceptibility to premature aging of the immune system. The phenomenon of “inflammaging” has begun to be evident in the last decades, as a consequence of increased life expectancy due to the introduction of cART. Quality of life and survival have improved substantially; however, PLWH are predisposed to chronic inflammatory conditions leading to age-associated diseases, such as inflammatory bowel disease, neurocognitive disorders, cardiovascular diseases, metabolic syndrome, bone abnormalities, and non-HIV-associated cancers. Several approaches have been studied in numerous uncontrolled and/or randomized clinical trials with the aim of reducing immune activation/inflammatory status in PLWH, none of which have achieved consistent results

CMV load quantitation by the automated VERSANT k-PCR Molecular System in plasma of transplanted patients

Background: Promptness of results, sensibility and multi-specimen processing represent a significant challenge for the molecular routine in a clinical virology laboratory; this is especially important for viral load in transplanted patients where a rapid diagnosis is essential to start a proper therapy. This study has been initially planned to establish the performance of the Automated VERSANT kPCR Molecular System for herpesviruses and adenovirus viral load quantification in plasma specimens from transplanted patients. Methods: So far, 100 plasma samples randomly collected from patients serially admitted to Sapienza University Hospital “Policlinico Umberto I” were examined. The VERSANT kPCR Molecular System with new VERSANT MiPLX Software Solution (Siemens Healthcare Diagnostics, Tarrytown, NY, USA) was used to extract DNA from samples. CMV amplification, detection and quantification were performed automatically on the AD module of the VERSANT kPCR Molecular System, adding quantitative standards in each run. Data obtained were compared with those from a currently used semi-automated system including EasyMAG (Biomerieux, Italia S.p.A.) extraction and Elite-MGB (ELITech Group S.p.A. Molecular Diagnostics, Italy) Real time PCR Amplification. Results: The VERSANT kPCR Molecular System with new VERSANT MiPLX Software Solution allowed to simultaneously process more than 90 plasma clinical samples in less than 3 hours. Detectable levels of CMV DNA were found in 22 out of 100 samples and a good correlation was observed by data obtained from the two assays used (r = 0.96 p < 0.0001). Sixty-two samples showed undetectable levels of CMV DNA using both methods; 16 specimens showed viremia values detectable but <250 copies/ml using ELITech System; 7 of these were quantified using VERSANT kPCR. The study is still in progress to increase the number of samples and to extend the analysis to other viruses clinically significant in transplanted patients. Conclusion: At the moment the analysis of the data indicates that: in plasma samples, a good correlation was observed when data are compared to those obtained by ELITech system; the new VERSANT MiPLX Software Solution proved to be flexible in his ability to combine extraction and amplification

Detection of SARS-CoV-2 RNA and antibodies in breast milk of infected mothers

The SARS-CoV-2 outbreak in December 2019 brought many challenges to be addressed. One concerns the possible transmission of the virus and protective antibodies against SARS-CoV-2 to newborns through breastfeeding. The aim of this study was the detection of SARS-CoV-2 RNA and antibodies in the milk of SARS-CoV-2 positive mothers. Milk and blood samples were collected from twelve women with SARS-CoV-2 positive nasopharyngeal swabs. Viral RNA was investigated by RT-PCR, and the presence of IgA, IgM, and IgG anti-SARS-CoV-2 was evaluated in both breast milk and maternal blood. All milk samples showed negative results for SARS-CoV-2 RNA. Eight women (66%) had a detectable level of anti -SARS-CoV-2 IgA in their milk. Of this group, only one sample presented simultaneously serum antiviral IgM and IgG while other three samples showed only anti-SARS-CoV-2 IgG. The remaining four mothers with anti-SARS-CoV-2 IgA in their breast milk had no serum antibodies against SARS-CoV-2. Finally, four mothers (34%) did not have any anti-SARS-CoV-2 antibodies in breast milk and serum, except one mother who had antiviral IgG and IgA in serum. Our results suggest that breastfeeding of SARS-CoV-2 infected mothers is safe and should be encouraged as breast milk transmits maternal antiviral antibodies which protect the infant while its immune system is immature

COVID-19; RT-PCR; SARS-CoV-2; seminal fluid; spermatozoa

Great concerns have been raised on SARS-CoV-2 impact on men's andrological well-being and one of the critically unanswered questions is whether it is present or not in the seminal fluid of infected subjects. The expression of ACE2 and TMPRSS2 in the testis and in the male genital tract allows speculations about a possible testicular involvement during the infection, possibly mediated by local and/or systemic inflammation that might allow a high viral load to overcome the haemato-testicular barrier. To date, few investigations have been carried out to ascertain the presence of SARS-CoV-2 in the seminal fluid with contrasting results. Furthermore, the cumulative number of subjects is far too low to answer the question unambiguously. Therefore, great caution is still needed when evaluating this data, otherwise we risk unleashing unmotivated concerns in the scientific world with troublesome consequences in reproductive medicine

Evaluation of performances of VERSANT HCV RNA 1.0 assay (kPCR) and Roche COBAS AmpliPrep/COBAS TaqMan HCV test v2.0 at low level viremia

We assess the concordance between low level HCV values obtained using the VERSANT HCV RNA 1.0 Assay (kPCR) and COBAS AmpliPrep/COBAS TaqMan HCV Quantitative Test v2.0. The correlation between the values obtained by the two RT-PCR assays for samples with quantifiable HCV RNA levels revealed that viral load measured by kPCR significantly correlated with that of the CAP/CTM (R=0.644, P<0.0001). The results show a good concordance (n=126/144, 87%); discordant results were mainly observed in the assessment of values below the lower limit of detection of the assays. These variations may have an impact on clinical decisions for patients on HCV triple therapy or interferon- free regimens. It is therefore recommended to monitor individual patients with the same test throughout treatment