Rapid diagnosis of bloodstream infections in the critically ill: Evaluation of the broad-range PCR/ESI-MS technology.

Bloodstream infection (BSI) and associated sepsis represent a major source of mortality in industrialized countries. Prompt treatment with targeted antibiotics affects both the financial impact and the clinical outcome of BSI: every hour gained in initiating the correct antimicrobial therapy significantly increases the probability of patient survival. However, the current standard-of-care, which depends on blood culture-based diagnosis, are often unable to provide such a fast response. Fast and sensitive molecular techniques for the detection of sepsis-related pathogens from primary blood samples are strongly needed. The aim of this study was to assess the usefulness of the IRIDICA BAC BSI Assay, a PCR/ESI-MS-based technology for the early diagnosis of bloodstream infections from primary blood samples in critical patients. This evaluation has been performed by comparison with the traditional culture-based methods. The study was performed on a total of 300 prospective whole blood specimens obtained from patients suspected of sepsis, admitted to enrolling ER units from The Greater Romagna Area. The overall concordance between the two techniques was of 86%, with a calculated sensitivity of 76% and an assay specificity of 90%. The clinical significance of discrepant results was evaluated reviewing the patients' clinical records and the results of additional relevant microbiological tests. The data here obtained support the ability of the IRIDICA BAC BSI Assay to identify a broad range of bacteria directly from primary whole blood samples, within eight hours. This might allow a timely administration of a suitable treatment

Serological and molecular tools to diagnose visceral leishmaniasis: 2-years' experience of a single center in Northern Italy

The diagnosis of visceral leishmaniasis (VL) remains challenging, due to the limited sensitivity of microscopy, the poor performance of serological methods in immunocompromised patients and the lack of standardization of molecular tests. The aim of this study was to implement a combined diagnostic workflow by integrating serological and molecular tests with standardized clinical criteria. Between July 2013 and June 2015, the proposed workflow was applied to specimens obtained from 94 in-patients with clinical suspicion of VL in the Emilia-Romagna region, Northern Italy. Serological tests and molecular techniques were employed. Twenty-one adult patients (22%) had a confirmed diagnosis of VL by clinical criteria, serology and/or real-time polymerase chain reaction; 4 of these patients were HIV-positive. Molecular tests exhibited higher sensitivity than serological tests for the diagnosis of VL. In our experience, the rK39 immunochromatographic test was insufficiently sensitive for use as a screening test for the diagnosis of VL caused by L. infantum in Italy. However, as molecular tests are yet not standardized, further studies are required to identify an optimal screening test for Mediterranean VL

IRIDICA-positive and blood culture-negative detections.

<p>IRIDICA-positive and blood culture-negative detections.</p

Unmatching results between the two methods.

<p>Unmatching results between the two methods.</p

Distribution of the organisms included in the study.

<p>The organisms reported by culture (solid bar) and PCR/ESI-MS (patterned bar) are sorted by decreasing order of PCR/ESI-MS reported organisms.</p

Concordant primary pathogen identification with unmatched additional detections.

<p>Concordant primary pathogen identification with unmatched additional detections.</p

Assay performance.

<p>Assay performance.</p

IRIDICA-negative and blood culture-positive detections.

<p>IRIDICA-negative and blood culture-positive detections.</p

Prevalence of HDV infection in Central Italy has remained stable across the last two decades with dominance of sub-genotypes 1 and characterized by elevated viral replication

Background: HDV-prevalence in Italy and its fluctuations over-time are controversial. Furthermore, an extensive characterization of HDV-infected patients is still missing. Methods: The rate of HDV-seroprevalence and HDV-chronicity was assessed in 1,579 HBsAg+patients collected from 2005 to 2022 in Central-Italy. Results: 45.3% of HBsAg+patients received HDV-screening with an increasing temporal-trend: 15.6% (2005-2010), 45.0% (2011-2014), 49.4% (2015-2018), 71.8% (2019-2022). By multivariable-model, factors correlated with the lack of HDV-screening were ALT&lt;2ULN and previous time-windows (P&lt;0.002). Furthermore, 13.4% of HDV-screened patients resulted anti-HDV+ with a stable temporal trend. Among them, 80.8% had detectable HDV-RNA (median[IQR]:4.6[3.6-5.6]logcopies/ml) with altered ALT in 89.3% (median[IQR]:92[62-177]U/L). Anti-HDV+ patients from Eastern/South-eastern Europe were younger than Italians (44[37-54] vs 53[47-62]years, P&lt;0.0001), less frequently NUC-treated (58.5% vs 80%, P=0.026) with higher HDV-RNA (4.8[3.6-5.8] vs 3.9[1.4-4.9]logcopies/ml, P=0.016) and HBsAg (9,461[4,159-24,532] vs 4,447[737-13,336]IU/ml, P=0.032). Phylogenetic-analysis revealed the circulation of HDV subgenotype-1e (47.4%) and -1c (52.6%). Notably, subgenotype-1e correlated with higher ALT than 1c (168[89-190] vs 58[54-88]U/l, P=0.015) despite comparable HDV-RNA. Conclusion: HDV-screening awareness is increasing over-time even if some gaps persist to achieve HDV-screening in all HBsAg+patients. HDV prevalence in tertiary-care centers tends to scarcely decline in native/non-native patients. Detection of subgenotypes, triggering variable inflammatory stimuli, supports the need to expand HDV molecular characterization