Microbiology and Virology Unit

Padova, Italy

Microbiology and Virology Unit

Padova, Italy
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Foti C.,University of Bari | Romita P.,University of Bari | Mascia P.,University of Bari | Miragliotta G.,Microbiology and Virology Unit | Calvario A.,Microbiology and Virology Unit
Endocrine, Metabolic and Immune Disorders - Drug Targets | Year: 2017

Context: Herpetic whitlow is caused by herpes virus (type1 or 2) during primary infection or as result of autoinoculation. Commonly, it is caused by HSV-2 in adults with positive history for genital infection. Case Description: We report the case of a 44-year-old woman that came to our attention with a 3- year history of recurrent cutaneous eruption on the ring finger of her left hand associated to lymphangitis of the homolateral arm. Laboratory exams including PCR on blood and cutaneous swab allowed to diagnosis it as a rare case of herpetic whitlow. Conclusion: The case here reported demonstrates that herpetic whitlow should be kept in mind by physicians in recurrent cases of fingers infection. Advanced diagnostic techniques as PCR are required to help clinicians to achieve a definite diagnosis and to choose the right treatment. © 2017 Bentham Science Publishers.

Franchin E.,Microbiology and Virology Unit | Barzon L.,Microbiology and Virology Unit | Cavallaro A.,Microbiology and Virology Unit | Richter S.N.,Microbiology and Virology Unit | Palua G.,Microbiology and Virology Unit
Journal of Clinical Microbiology | Year: 2012

An extremely drug-resistant Klebsiella pneumoniae isolate, sequence type ST101, was isolated from a patient in Italy. We describe antibiotic treatment, measures to clear and contain the infection, and a complete sequence analysis of a novel large plasmid, pKPN101-IT, harboring the blaKPC-2 gene and arising from the threatening recombination of different worldwide-distributed backbones. Copyright © 2012, American Society for Microbiology. All Rights Reserved.

Richter S.N.,Microbiology and Virology Unit | Biasolo M.A.,Microbiology and Virology Unit | Bartolini A.,Microbiology and Virology Unit | Cavallaro A.,Microbiology and Virology Unit | Palu G.,Microbiology and Virology Unit
Journal of Clinical Microbiology | Year: 2012

The novel real-time PCR assay developed as described here was able to detect bla KPC1/2-12 (bla KPC-1/2 to bla KPC-12) from easily available clinical specimens in less than 2 h. The genotypic assay was highly sensitive (100%) and specific (98%). In some cases, it was able to detect bla KPC 48 h before positive detection by standard phenotypic assay on patients who were monitored daily. The high sensitivity and rapidity of the molecular method make it the method of choice for KPC surveillance and, thus, containment purposes. Copyright © 2012, American Society for Microbiology. All Rights Reserved.

PubMed | Microbiology and Virology Unit and Amedeo Of Savoia Hospital
Type: Journal Article | Journal: The new microbiologica | Year: 2016

In immunosuppressed patients, pre-emptive therapy and a strict follow-up of CMV infection are the standard of care for the prevention of CMV disease. Several real-time PCR assays for CMV DNA quantification on whole blood (WB) and plasma (PL) are commercially available. This study compared and correlated CMV viral loads obtained by the Cobas AmpliPrep/Cobas TaqMan (CAP/CTM) platform on plasma specimens with those obtained on corresponding whole blood specimens by the real-time PCR assay (ELITe MGB-CMV) in 185 sequential samples from 41 immunosuppressed patients. Correlation between the two assays was good. Kinetics of CMV DNA within the same patient was similar, but PL viral load was constantly 1 log lower than WB. In patients under antiviral therapy, low level of CMV DNA persisted in WB, while it was absent in PL. The good correlation between CMV DNA detected on both PL and WB supports the reliability of the two matrices for viral monitoring and the therapeutic management of CMV infection. Nevertheless, due to significant quantification differences between PL and WB CMV DNA, the same biological specimen should be used for a sequential and reliable follow-up of patients at high risk of CMV infection.

PubMed | University of Verona, San Raffaele Scientific Institute, Cystic Fibrosis Center and Microbiology and Virology Unit
Type: Journal Article | Journal: BMC microbiology | Year: 2016

Pseudomonas aeruginosa is the predominant pathogen associated with the decline of pulmonary function in cystic fibrosis (CF) patients. Both environment-to-host acquisition and patient-to-patient transmission have been described for P. aeruginosa infection. Epidemic clones and bacterial phenotypic adaptation to the CF lung have been recognised as independent risk factors for disease progression. So far, there is no established link between genotypic prevalence and phenotypic traits. Here, we look at the major CF patient cohort in Italy to identify shared P. aeruginosa clones and associated common phenotypic traits.A comprehensive analysis of P. aeruginosa genotypes to determine the presence of high-risk shared clones and their association to specific phenotypic traits has been performed in a major Italian CF centre. Pulsed-field gel electrophoresis (PFGE) of P. aeruginosa isolates from 338 CF subjects identified 43 profiles shared by two or more patients and 214 profiles exclusive to individual patients. There was no evidence of a P. aeruginosa outbreak, but four most prevalent pulsotypes were detected. Common phenotypic traits were recorded intra-pulsotypes, but we detected heterogeneity inter-pulsotypes. Two of the four major pulsotypes included P. aeruginosa isolates with hallmarks of adaptation to the CF airways, including loss of motility, low production of siderophore, pyocyanin and proteases, and antibiotic resistance. One of these pulsotypes grouped a high percentage of hypermutable isolates. No clear correlation between epidemiological and clinical data was found.We conclude that CF patients of this cohort shared common pulsotypes, but their phenotypic heterogeneity indicates an absence of specific traits associated to P. aeruginosa genotypic prevalence.

PubMed | University of Turin and Microbiology and Virology Unit
Type: | Journal: Current microbiology | Year: 2016

Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a useful tool for rapid identification of microorganisms. Unfortunately, its direct application to positive blood culture is still lacking standardized procedures. In this study, we evaluated an easy- and rapid-to-perform protocol for MALDI-TOF MS direct identification of microorganisms from positive blood culture after a short-term incubation on solid medium. This protocol was used to evaluate direct identification of microorganisms from 162 positive monomicrobial blood cultures; at different incubation times (3, 5, 24h), MALDI-TOF MS assay was performed from the growing microorganism patina. Overall, MALDI-TOF MS concordance with conventional methods at species level was 60.5, 80.2, and 93.8% at 3, 5, and 24h, respectively. Considering only bacteria, the identification performances at species level were 64.1, 85.0, and 94.1% at 3, 5, and 24h, respectively. This protocol applied to a commercially available MS typing system may represent, a fast and powerful diagnostic tool for pathogen direct identification and for a promptly and pathogen-driven antimicrobial therapy in selected cases.

Frasson I.,University of Padua | Biasolo M.A.,University of Padua | Biasolo M.A.,Microbiology and Virology Unit | Bartolini A.,Microbiology and Virology Unit | And 5 more authors.
International Journal of Antimicrobial Agents | Year: 2013

VIM and KPC are two major families of carbapenemases involved in nosocomial outbreaks of multidrug-resistant Gram-negative bacilli. To rapidly detect blaVIM- and blaKPC-encoding strains, three multiplex PCR-based methods were designed and validated: (i) a real-time PCR to detect all reported VIM alleles, namely blaVIM-1-19, 23-37; (ii) a real-time PCR to identify blaVIM-type and blaKPC carbapenemases in an ultrarapid single reaction; and (iii) a standard PCR to amplify and sequence all VIM alleles. All three methods detected 33 VIM-positive samples among 107 Gram-negative isolates with imipenem and meropenem minimum inhibitory concentrations ≥1 mg/L. The three methods displayed 100% sensitivity, specificity and concordance. Sequencing of the blaVIM amplicons revealed that 30 samples encoded blaVIM-1 and 3 samples encoded blaVIM-2. The real-time assay, optimised for the simultaneous detection of blaVIM and blaKPC, identified 3 and 12 isolates positive for both blaVIM/blaKPC and for bla KPC, respectively. The analytical sensitivity of the real-time assays was linear over 6 log dilutions, with a reproducible detection limit of 1 CFU. No cross-reactivity was detected. The developed assays provide powerful tools for rapid identification of VIM and KPC carbapenemase producers, therefore contributing to the prevention and containment of resistance dissemination. © 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

Richter S.N.,University of Padua | Richter S.N.,Microbiology and Virology Unit | Frasson I.,University of Padua | Bergo C.,Microbiology and Virology Unit | And 5 more authors.
Journal of Clinical Microbiology | Year: 2011

The first case in Europe of Klebsiella pneumoniae carbapenemase (KPC) 2 transfer from K. pneumoniae to Escherichia coli in the same patient is described. KPC-positive plasmids from the two species were identical, indicating horizontal plasmid transfer. Selection of the KPC-producing E. coli strain was triggered by therapy with meropenem. Copyright © 2011, American Society for Microbiology. All Rights Reserved.

Richter S.N.,University of Padua | Richter S.N.,Microbiology and Virology Unit | Frasson I.,University of Padua | Franchin E.,University of Padua | And 8 more authors.
Gut Pathogens | Year: 2012

Background: Klebsiella pneumoniae carbapenemases (KPCs) producing bacteria have emerged as a cause of multidrug-resistant nosocomial infections worldwide. KPCs are plasmid-encoded enzymes capable of hydrolysing a broad spectrum of beta-lactams, including carbapenems and monobactams, therefore worryingly limiting antimicrobial treatment options. Analysis of circulating bacterial strains and KPC alleles may help understanding the route of KPC dissemination and therefore help containing the infection. Methods: KPC-producing Klebsiella pneumoniae dissemination in two 1580- and 300- bed hospitals in Padua, Italy, from initial outbreak in 2009 to late 2011 was analysed. Molecular and clinical epidemiology, including bacterial strains, KPC-encoding plasmid sequences and associated resistance genes, involved hospital wards and relocation of patients were described. Routine antimicrobial susceptibility testing and MIC of carbapenems on clinical isolates were performed. Detection of resistance genes was obtained by PCR and sequencing. MLST, PFGE and ERIC were used for molecular genotyping. Plasmid analysis was obtained by digestion with restriction enzymes and deep sequencing. Results: KPC-positive clinical samples were isolated from nearly 200 patients. In the initial outbreak intensive care units were almost exclusively involved, while medical, surgical and long-term wards were successively massively concerned. Analysis of KPC alleles, plasmids and bacterial sequence types (STs) indicated that during the initial outbreak KPC-3 in ST258 and KPC-2 in ST147 were each confined in one of the two surveilled hospitals. While KPC-2 dissemination was effectively contained, KPC-3 in ST258 cross-spreading was observed. The simultaneous presence of two carbapenemases, VIM-1 and KPC-2, in the same isolate was also observed in three patients. Total sequencing of plasmid content of two KPC-3 strains showed novel association of resistance plasmids. Conclusions: The acquired molecular epidemiology demonstrated that 1) both acquisitions from outward sources and patient relocation within the hospitals were responsible for the observed spreading; 2) KPC-3-encoding Klebsiella pneumoniae ST258 prevailed over other strains. In addition, the described massive transfer of KPC-mediated resistance to non-intensive care units may anticipate spreading of resistance to the non-hospitalized population. Therefore, genotypic analysis alongside phenotypic identification of carbapenemase producers, also at the carriage state, is advisable to prevent and contain further carbapenemase resistance dissemination. © 2012 Richter et al.; licensee BioMed Central Ltd.

Costa C.,Microbiology and Virology Unit | Balloco C.,Microbiology and Virology Unit | Sidoti F.,Microbiology and Virology Unit | Mantovani S.,Microbiology and Virology Unit | And 5 more authors.
Journal of Clinical Virology | Year: 2014

Background: Immunological monitoring for CMV can be useful in transplant patients; however, few centers perform it on a routine basis. Objectives: In this study, CMV-specific cellular response was evaluated in a population of kidney transplant recipients and related to viral infection/reactivation and other demographic and clinical features. Study design: Three hundred and twenty-eight patients were studied by EliSPOT assay: 201 prospectively monitored in the first year posttransplantation, 127 with a single determination at >1 year. Clinical features, including occurrence of CMV-DNAemia, CMV serostatus, anti-viral strategies and immunosuppressive protocols, were evaluated. Results: Overall, 66.5% of patients were CMV-responders at EliSPOT assay. No episode of infection occurred at follow-up (mean 24.5 months) in 73.4% responders versus 55.5% non-responders (p<0.005); CMV-free period was significantly longer in responders (p<0.001). Although no significant difference of peak viral load was found, prevalence of CMV-DNAemia values >105copies/mL was significantly higher in non-responders versus responders (8.2% and 2.3%, p<0.05). Non-responder status was significantly associated to CMV-seronegativity (p<0.0001), anti-viral prophylaxis use (p<0.0001), and immunosuppression induction with basiliximab (p<0.005). No significant association was found for other clinical features and immunosuppressive protocols. Conclusions: Immunological data for CMV could be used in the clinical evaluation and decision-making process, in combination with virological monitoring, in kidney transplant recipients. © 2014 Elsevier B.V.

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