SOSTIENICI
Zannoli Dott.ssa Silvia
Pubblicazioni su PubMed
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Correlating qRT-PCR, dPCR and Viral Titration for the Identification and Quantification of SARS-CoV-2: A New Approach for Infection Management.
Viruses2021 05;13(6):. doi: 1022.
Brandolini Martina, Taddei Francesca, Marino Maria Michela, Grumiro Laura, Scalcione Agata, Turba Maria Elena, Gentilini Fabio, Fantini Michela, Zannoli Silvia, Dirani Giorgio, Sambri Vittorio
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in Wuhan, China, in late 2019 and is the causative agent of the coronavirus disease 2019 (COVID-19) pandemic. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) represents the gold standard for diagnostic assays even if it cannot precisely quantify viral RNA copies. Thus, we decided to compare qRT-PCR with digital polymerase chain reaction (dPCR), which is able to give an accurate number of RNA copies that can be found in a specimen. However, the aforementioned methods are not capable to discriminate if the detected RNA is infectious or not. For this purpose, it is necessary to perform an endpoint titration on cell cultures, which is largely used in the research field and provides a tissue culture infecting dose per mL (TCID50/mL) value. Both research and diagnostics call for a model that allows the comparison between the results obtained employing different analytical methods. The aim of this study is to define a comparison among two qRT-PCR protocols (one with preliminary RNA extraction and purification and an extraction-free qRT-PCR), a dPCR and a titration on cell cultures. The resulting correlations yield a faithful estimation of the total number of RNA copies and of the infectious viral burden from a Ct value obtained with diagnostic routine tests. All these estimations take into consideration methodological errors linked to the qRT-PCR, dPCR and titration assays.
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Multicenter Evaluation of the C6 Lyme ELISA Kit for the Diagnosis of Lyme Disease.
Microorganisms2020 Mar;8(3):. doi: E457.
Zannoli Silvia, Fantini Michela, Semprini Simona, Marchini Barbara, Ceccarelli Barbara, Sparacino Monica, Schiavone Pasqua, Belgrano Anna, Ruscio Maurizio, Gobbetti Martina, Nicoletti Maira, Robatscher Eva, Pagani Elisabetta, Sambri Vittorio
Abstract
Lyme disease (LD), caused by infection with , is the most common tick-borne infection in many regions of Eurasia. Antibody detection is the most frequently used laboratory test, favoring a two-step serodiagnostic algorithm; immunoenzymatic detection of antibodies to C6 has been shown to perform similarly to a standard two-step workflow. The aim of this study was the performance evaluation of the C6 Lyme ELISA kit compared to a standard two-step algorithm in three laboratories located in the northeastern region of Italy which cater to areas with different LD epidemiology. A total of 804 samples were tested, of which 695 gave concordant results between C6 testing and routine workflow (564 negative, 131 positive). Wherever available, clinical presentation and additional laboratory tests were analyzed to solve discrepancies. The C6 based method showed a good concordance with the standard two-step algorithm (Cohen's ? = 0.619), however, the distribution of discrepancies seems to point towards a slightly lower specificity of C6 testing, which is supported by literature and could impact on patient management. The C6 ELISA, therefore, is not an ideal stand-alone test; however, if integrated into a two-step algorithm, it might play a part in achieving a sensitive, specific laboratory diagnosis of LD.
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Diagnosing Clostridioides difficile infections with molecular diagnostics: multicenter evaluation of revogene C. difficile assay.
Eur J Clin Microbiol Infect Dis2020 Jun;39(6):1169-1175. doi: 10.1007/s10096-020-03829-4.
Sambri Vittorio, Gateau Cécile, Zannoli Silvia, Dirani Giorgio, Couturier Jeanne, Op den Buijs Ingrid, Roymans René, Hallet Emma, Arnold Mihaela, Zumoberhaus Anja, Steiner Stanislava, van de Bovenkamp Jeroen, Altwegg Martin, Berlinger Livia, Barbut Frédéric,
Abstract
Clostridioides difficile infections are a significant threat to our healthcare system, and rapid and accurate diagnostics are crucial to implement the necessary infection prevention and control measurements. Nucleic acid amplification tests are such reliable diagnostic tools for the detection of toxigenic Clostridioides difficile strains directly from stool specimens. In this multicenter evaluation, we determined the performance of the revogene C. difficile assay. The analysis was conducted on prospective stool specimens collected from six different sites in Europe. The performance of the revogene C. difficile assay was compared to the different routine diagnostic methods and, for a subset of the specimens, against toxigenic culture. In total, 2621 valid stool specimens were tested, and the revogene C. difficile assay displayed a sensitivity/specificity of 97.1% [93.3-99.0] and 98.9% [98.5-99.3] for identification of Clostridioides difficile infection. Discrepancy analysis using additional methods improved this performance to 98.8% [95.8-99.9] and 99.6% [99.2-99.8], respectively. In comparison to toxigenic culture, the revogene C. difficile assay displayed a sensitivity/specificity of 93.0% [86.1-97.1] and 99.5% [98.7-99.9], respectively. These results indicate that the revogene C. difficile assay is a robust and reliable aid in the diagnosis of Clostridioides difficile infections.
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Comparison of Four Commercial Screening Assays for the Detection of bla, bla, bla, bla and bla in Rectal Secretion Collected by Swabs.
Microorganisms2019 Dec;7(12):. doi: E704.
Del Bianco Francesca, Morotti Manuela, Zannoli Silvia, Dirani Giorgio, Fantini Michela, Pedna Maria Federica, Farabegoli Patrizia, Sambri Vittorio
Abstract
The spread of carbapenem-resistant Enterobacteriaceae (CRE) has been enabled by the lack of control measures directed at carriers of multidrug-resistant organisms in healthcare settings. Screening patients for asymptomatic colonization on the one hand, and implementation of contact precautions on the other hand, reduces patient-to-patient transmission. Screening plates represents a relatively low-cost method for isolating CRE from rectal swabs; however, molecular assays have become widely available. This study compared the performance of four commercial molecular platforms in detecting clinically significant carbapenemase genes versus routine screening for CRE. A total of 1015 non-duplicated rectal swabs were cultured on a chromogenic carbapenem-resistant selective medium. All growing Enterobacteriaceae strains were tested for carbapenemase-related genes. The same specimens were processed using the following molecular assays: Allplex? Entero-DR, Amplidiag CarbaR + MCR, AusDiagnostics MT CRE EU, and EasyScreen? ESBL/CPO. The prevalence of carbapenemase (KPC)-producing Enterobacteriaceae detected by swab culture was 2.2%, while organisms producing oxacillinase (OXA)-48 and metallo-?-lactamases were infrequent. The cost of CRE-related infection control precautions, which must be kept in place while waiting for screening results, are significant, so the molecular tests could become cost-competitive, especially when the turnaround time is decreased dramatically. Molecular assays represent a powerful diagnostic tool as they allow the rapid detection of the most clinically relevant carbapenemases.
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Unyvero ITI system for the clinical resolution of discrepancies in periprosthetic joint infection diagnosis.
Musculoskelet Surg2021 Apr;105(1):39-42. doi: 10.1007/s12306-019-00626-x.
Zannoli Silvia, Sambri Andrea, Morotti Manuela, Tassinari Martina, Torri Arianna, Bianchi Giuseppe, De Paolis Massimiliano, Donati Davide Maria, Sambri Vittorio
Abstract
OBJECTIVES:
The Unyvero molecular assay was tested for the clinical resolution of discordant results, evaluating its role in prosthetic joint infection diagnosis.
METHODS:
Multiplex PCR was performed on 45 samples from prosthesis treatment (either sonication or dithiothreitol). Analytical performance was compared to that of biofilm culture using Musculoskeletal Infection Society criteria as gold standard.
RESULTS:
Unyvero and biofilm culture showed similar agreement rates compared to the gold standard (34/43 and 32/43, respectively). Both methods showed six additional identifications compatible with true infection; five positive results from biofilm culture were deemed contaminations.
CONCLUSIONS:
The Unyvero system showed good performances and a significantly shorter turnaround time compared to cultural methods, presenting an added value to PJI diagnosis even when performed following a composite approach.
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West Nile Virus and Usutu Virus Co-Circulation in Europe: Epidemiology and Implications.
Microorganisms2019 Jun;7(7):. doi: E184.
Zannoli Silvia, Sambri Vittorio
Abstract
West Nile virus (WNV) and Usutu virus (USUV) are neurotropic mosquito-borne flaviviruses that may infect humans. Although WNV is much more widespread and plays a much larger role in human health, the two viruses are characterized by similar envelope antigens, clinical manifestations, and present overlapping in terms of geographic range of transmission, host, and vector species. This review highlights some of the most relevant aspects of WNV and USUV human infections in Europe, and the possible implications of their co-circulation.
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Easyscreen? Enteric Protozoa Assay For the Detection of Intestinal Parasites: A Retrospective Bi-Center Study.
J Parasitol2019 02;105(1):58-63.
Dirani Giorgio, Zannoli Silvia, Paesini Elena, Farabegoli Patrizia, Dalmo Barbara, Vocale Caterina, Liguori Giovanna, Varani Stefania, Sambri Vittorio
Abstract
Gastroenteritis caused by single or multiple pathogens remains a major diagnostic challenge for the laboratory, as diagnosis is achieved using different techniques with variable sensitivity and specificity. The aim of this study was to evaluate the EasyScreen? Enteric Protozoa Detection Kit, a multiplex PCR assay for the detection and identification of the 5 most common protozoan parasites in fecal samples. A total of 632 fecal samples, submitted for routine screening to 2 centers in north-eastern Italy, was included in the study. The results of the molecular assay were compared to those of the standard diagnostic procedures, represented by microscopy and immunoassays. Out of 32 samples testing positive by conventional tools, 31 were detected as concordantly positive using the EasyScreen Kit. Additionally, 91 out of 632 samples only tested positive by the molecular test, therefore increasing the positive detection rate by 275%. Finally, the EasyScreen assay detected 14 co-infections compared to 3 co-infections identified by conventional methods. The EasyScreen Kit provided a rapid and sensitive simultaneous identification of the most important diarrhea-causing protozoa that infect humans. Additionally, this molecular assay presents several advantages compared to conventional tools, such as the standardization and near-total automation of the process. Although critical issues related to the employment of molecular assays are still evident, the system is suitable for clinical parasitological diagnosis as long as it is used in association with conventional tools.
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Rapid diagnosis of bloodstream infections in the critically ill: Evaluation of the broad-range PCR/ESI-MS technology.
PLoS One2018 ;13(5):e0197436. doi: 10.1371/journal.pone.0197436.
Tassinari Martina, Zannoli Silvia, Farabegoli Patrizia, Pedna Maria Federica, Pierro Anna, Mastroianni Antonio, Fontan Riccardo, Luongo Luciano, Sarnataro Giuseppe, Menegatti Elisa, Caruso Assunta, Sambri Vittorio
Abstract
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.
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Is Treatment With Dithiothreitol More Effective Than Sonication for the Diagnosis of Prosthetic Joint Infection?
Clin Orthop Relat Res2018 01;476(1):137-145. doi: 10.1007/s11999.0000000000000060.
Sambri Andrea, Cadossi Matteo, Giannini Sandro, Pignatti Giovanni, Marcacci Maurilio, Neri Maria Pia, Maso Alessandra, Storni Elisa, Gamberini Simonetta, Naldi Susanna, Torri Arianna, Zannoli Silvia, Tassinari Martina, Fantini Michela, Bianchi Giuseppe, Donati Davide, Sambri Vittorio
Abstract
BACKGROUND:
Prosthetic joint infection (PJI) is among the most-severe complications of a total joint arthroplasty. Identification of the causal organism is of paramount importance for successful treatment, and sonication of implants may aid in this identification. Dithiothreitol (DTT) treatment has been proposed as an alternative to sonication to improve diagnosis, reduce costs, and improve reliability of the procedure, but its efficacy remains poorly characterized.
QUESTIONS/PURPOSES:
(1) Are DTT and sonication more sensitive and/or more specific than standard cultures of tissue samples for the diagnosis of PJI? (2) Which test (DTT or sonication) is more sensitive when the clinician does not suspect infection before surgery? (3) Which test (DTT or sonication) is more sensitive when the clinician suspects infection before surgery?
METHODS:
Two hundred thirty-two patients undergoing revision of a knee or hip arthroplasty were prospectively evaluated in this randomized study. Cultures were performed on five tissue samples from each patient and on fluid obtained by prosthesis treatment in patients randomly assigned to sonication (117 patients) or DTT (115 patients). The reference standard against which cultures (on tissue samples and on fluids from sonication or DTT) were compared was the Musculoskeletal Infection Society definition of PJI.
RESULTS:
Cultures on sonication and DTT fluids provided higher sensitivity (89% and 91%, respectively) than those on standard cultures of tissue samples (79%; p
CONCLUSIONS:
In this randomized study, we found no difference in sensitivity between DTT and sonication for the detection of PJI, and both of those tests were more sensitive than standard tissue cultures. Thus, cultures of sonication or DTT fluid should be considered important additional tools to standard cultures for definition of PJI and should be considered together with other criteria, especially in settings where infection is not suspected before revision surgery.Level of Evidence Level I, diagnostic study.
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Pulmonary function and expiratory flow limitation in acute cervical spinal cord injury.
Arch Phys Med Rehabil2012 Nov;93(11):1950-6. doi: 10.1016/j.apmr.2012.04.015.
Alvisi Valentina, Marangoni Elisabetta, Zannoli Silvia, Uneddu Mariella, Uggento Riccardo, Farabegoli Lucia, Ragazzi Riccardo, Milic-Emili Joseph, Belloni Gian P, Alvisi Raffaele, Volta Carlo A
Abstract
OBJECTIVE:
To identify the nature of the changes of respiratory mechanics in patients with middle cervical spinal cord injury (SCI) and their correlation with posture.
DESIGN:
Clinical trial.
SETTING:
Acute SCI unit.
PARTICIPANTS:
Patients with SCI (N=34) at C4-5 level studied within 6 months of injury.
INTERVENTIONS:
Patients were assessed by the negative expiratory pressure test, maximal static respiratory pressure test, and standard spirometry.
MAIN OUTCOME MEASURES:
The following respiratory variables were recorded in both the semirecumbent and supine positions: (1) tidal expiratory flow limitation (TEFL); (2) airway resistances; (3) mouth occlusion pressure developed 0.1 seconds after occluded inspiration at functional residual capacity (P(0.1)); (4) maximal static inspiratory pressure (MIP) and maximal static expiratory pressure (MEP); and (5) spirometric data.
RESULTS:
TEFL was detected in 32% of the patients in the supine position and in 9% in the semirecumbent position. Airway resistances and P(0.1) were much higher compared with normative values, while MIP and MEP were markedly reduced. The ratio of forced expiratory volume in 1 second to forced vital capacity was less than 70%, while the other spirometric data were reduced up to 30% of predicted values.
CONCLUSIONS:
Patients with middle cervical SCI can develop TEFL. The presence of TEFL, associated with increased airway resistance, could increase the work of breathing in the presence of a reduced capacity of the respiratory muscles to respond to the increased load. The semirecumbent position and the use of continuous positive airway pressure can be helpful to (1) reduce the extent of TEFL and avoid the opening/closure of the small airways; (2) decrease airway resistance; and (3) maintain the expiratory flow as high as possible, which aids in the removal of secretions.
Copyright © 2012 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.
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