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ABSTRACT: We evaluated a three-step algorithm for laboratory diagnosis of Clostridium difficile-associated diarrhoea (CDAD). First, stool specimens were screened using an EIA test for glutamate dehydrogenase detection. Screen-positive specimens were tested by a rapid cytotoxintoxin A/B assay and subjected to stool culture. All cultures positive for C. difficile underwent toxigenic culture. The results showed that toxigenic culture allowed us to recover 37/156 (24.4%) stool samples harbouring toxigenic C. difficile that would have been missed by using faecal cytotoxin assay alone. This determined an increase in infection prevalence of 4.2% (from 11.4% to 15.6 %). Furthermore, to characterize the clinical Clostridium difficile isolates and the distribution of PCR ribotypes circulating in the San Carlo Borromeo hospital, molecular typing using semi-automated repetitive-sequence-based PCR (rep- PCR) and PCR ribotyping, and an evaluation of the antibiotic resistance were also performed. Among them, 71 indistinguishable strains were detected by rep-PCR and 83 by PCR-ribotyping revealing C. difficile outbreaks in our hospital. A total of 6 different ribotypes were obtained by PCR ribotyping. The most frequent ribotype was 018 (88.2%) that also showed resistance to moxifloxacin. In one case, uncommon PCR ribotype 186 was also identified.
The new microbiologica: official journal of the Italian Society for Medical, Odontoiatric, and Clinical Microbiology (SIMMOC) 07/2012; 35(3):307-16. · 1.00 Impact Factor
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ABSTRACT: Fungaemia is an increasing nosocomial pathology. The 'gold standard' for detection of fungaemia is blood culture, but it is time-consuming and its sensitivity for early detection is low. On the other hand, yeasts present different antifungal sensitivity patterns to be quickly detected to allow an effective treatment. The aim of this study was to evaluate the diagnostic performances of PNA-FISH to directly identify yeasts from blood cultures and to compare results with those obtained by culture. A total of 176 blood cultures positive for yeasts at direct Gram stain and 24 negative blood cultures as control collected from 15 Italian hospitals, included in a network coordinated by the Medical Mycology Committee, Italian Society of Clinical Microbiology (AMCLI), were examined both by culture and PNA-FISH technology. Sensitivity of the PNA-FISH technique evaluated for five Candida species was 99.3% and specificity, 100%. Distinguishing which yeast is implicated in fungaemia and whether the infection is caused by multiple species are important for the selection of antifungal therapy. The PNA-FISH technique is a very useful approach because the test discriminates between groups of Candida species with different susceptibility pattern, particularly against azoles and echinocandins, with only a 90-minute turn-around time after the Gram-stain reading.
Mycoses 01/2012; 55(5):388-92. · 2.25 Impact Factor
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Claudio Farina,
Giuseppe Russello,
Stefano Andreoni,
Cristina Bonetti,
Marco Conte,
Paolo Fazi,
Gianluigi Lombardi,
Francesco Luzzaro,
Esther Manso,
Piero Marone,
Marco Passera,
Andrea Rocchetti,
Silvana Sanna,
Egidio Franco Viganò
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ABSTRACT: The authors evaluated the performance of the MycArray™ Yeast ID (Myconostica Ltd, UK) assay in the identification of a total of 88 yeast isolates recovered in culture as compared to that obtained through routine methods. The turn-around time for species identification directly from cultures by the MycArray was 6 hours, much quicker than classical methods and all yeasts were correctly identified. In two cases a double identification including Saccharomyces cerevisiae was noted, but it was not confirmed by culture. The results show that MycArray Yeast ID can be a potential tool for rapid detection and identification of Candida species.
Medical mycology: official publication of the International Society for Human and Animal Mycology 01/2012; 50(5):549-55. · 2.13 Impact Factor
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ABSTRACT: Many case reports suggest that the use of hyperbaric oxygen therapy associated with surgical and medical approaches may contribute to restricting the growth of zygomycetes in patient tissue. The primary aim of this study was to obtain data concerning the in vitro susceptibility of 22 zygomycetes to antifungals such as amphotericin B and posaconazole, and to compare the in vitro development of these fungi in aerobic normobaric versus hyperoxic normobaric and hyperbaric atmosphere. None of the zygomycetes grew after 24-hour and 72-hour incubation in a hyperoxic hyperbaric (2 or 3 ATA) atmosphere. However, when plates were maintained at room temperature in aerobic conditions, colonies were observed from 36-96 h after inoculation, while minimum inhibitory concentration (MIC) values remained the same. This preliminary in vitro study focuses on the in vitro examination of combination therapies to potentiate antifungal activity. Both hyperoxic hyperbaric conditions and a single antifungal agent, as well as combinations of different antifungal drugs were used. Results suggest an impressive in vitro fungistatic activity of the hyperoxic hyperbaric atmosphere, even if the antifungal effect is strictly time-dependent using these incubation conditions.
Medical mycology: official publication of the International Society for Human and Animal Mycology 09/2011; 50(4):427-32. · 2.13 Impact Factor
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ABSTRACT: An interlaboratory evaluation (seven centers) of VITEK2 System and Sensititre YeastOne® was conducted to test the antifungal susceptibilities of yeasts. The MICs of amphotericin B, fluconazole, flucytosine, and voriconazole were determined for 70 isolates of Candida spp. Our results demonstrated a higher interlaboratory agreement of VITEK 2 System than Sensititre YeastOne©. A good concordance between the two methods was observed for amphotericin B, fluconazole, voriconazole and 5-fluorocytosine (from 81.4% to 88.6%). The study suggests the potential value of the VITEK2 System as a convenient alternative method for testing the susceptibility of yeasts. It also indicates the need for further optimization of MIC endpoint criteria to improve interlaboratory agreement.
The new microbiologica: official journal of the Italian Society for Medical, Odontoiatric, and Clinical Microbiology (SIMMOC) 04/2011; 34(2):195-201. · 1.00 Impact Factor
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Alessandra Zappa,
Marta Canuti,
Elena Frati,
Elena Pariani,
Silvana Perin,
Maria Lorena Ruzza, Claudio Farina,
Alberto Podestà,
Alessandro Zanetti,
Antonella Amendola,
Elisabetta Tanzi
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ABSTRACT: The discovery of human Metapneumovirus (hMPV) and human Bocavirus (hBoV) identified the etiological causes of several cases of acute respiratory tract infections in children. This report describes the molecular epidemiology of hMPV and hBoV infections observed following viral surveillance of children hospitalized for acute respiratory tract infections in Milan, Italy. Pharyngeal swabs were collected from 240 children ≤3 years of age (130 males, 110 females; median age, 5.0 months; IQR, 2.0-12.5 months) and tested for respiratory viruses, including hMPV and hBoV, by molecular methods. hMPV-RNA and hBoV-DNA positive samples were characterized molecularly and a phylogenetical analysis was performed. PCR analysis identified 131/240 (54.6%) samples positive for at least one virus. The frequency of hMPV and hBoV infections was similar (8.3% and 12.1%, respectively). Both infections were associated with lower respiratory tract infections: hMPV was present as a single infectious agent in 7.2% of children with bronchiolitis, hBoV was associated with 18.5% of pediatric pneumonias and identified frequently as a single etiological agent. Genetically distinct hMPV and hBoV strains were identified in children examined with respiratory tract infections. Phylogenetic analysis showed an increased prevalence of hMPV genotype A (A2b sublineage) compared to genotype B (80% vs. 20%, respectively) and of the hBoV genotype St2 compared to genotype St1 (71.4% vs. 28.6%, respectively). Interestingly, a shift in hMPV infections resulting from A2 strains has been observed in recent years. In addition, the occurrence of recombination events between two hBoV strains with a breakpoint located in the VP1/VP2 region was identified.
Journal of Medical Virology 01/2011; 83(1):156-64. · 2.82 Impact Factor
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Giulia Morace,
Elisa Borghi,
Roberta Iatta,
Gerardino Amato,
Stefano Andreoni,
Gioconda Brigante, Claudio Farina,
Giuliana Lo Cascio,
Gianluigi Lombardi,
Ester Manso,
Michele Mussap,
Patrizia Pecile,
Roberto Rigoli,
Elisabetta Tangorra,
Maria Valmarin,
Maria Teresa Montagna
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ABSTRACT: Yeasts are a common cause of invasive fungal infections in critically ill patients. Antifungal susceptibility testing results of clinically significant fungal strains are of interest to physicians, enabling them to adopt appropriate strategies for empiric and prophylactic therapies. We investigated the antifungal susceptibility of yeasts isolated over a 2-year period from hospitalised patients with invasive yeast infections.
638 yeasts were isolated from the blood, central venous catheters and sterile fluids of 578 patients on general and surgical intensive care units and surgical wards. Etest strips and Sensititre panels were used to test the susceptibility of the isolates to amphotericin B, anidulafungin, caspofungin, fluconazole, itraconazole, posaconazole and voriconazole in 13 laboratories centres (LC) and two co-ordinating centres (CC). The Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution method was used at the CCs for comparison.
Etest and Sensititre (LC/CC) MIC90 values were, respectively: amphotericin B 0.5/0.38, 1/1 mg/L; anidulafungin 2/1.5 and 1/1 mg/L; caspofungin 1/0.75 and 0.5/0.5 mg/L; fluconazole 12/8 and 16/16 mg/L; itraconazole 1/1.5, 0.5/0.5 mg/L; posaconazole 0.5 mg/L and voriconazole 0.25 mg/L for all. The overall MIC90 values were influenced by the reduced susceptibility of Candida parapsilosis isolates to echinocandins and a reduced or lack of susceptibility of Candida glabrata and Candida krusei to azoles, in particular fluconazole and itraconazole. Comparison of the LC and CC results showed good Essential Agreement (90.3% for Etest and 92.9% for Sensititre), and even higher Categorical Agreement (93.9% for Etest and 96% for Sensititre); differences were observed according to the species, method, and antifungal drug. No cross-resistance between echinocandins and triazoles was detected.
Our data confirm the different antifungal susceptibility patterns among species, and highlight the need to perform antifungal susceptibility testing of clinically relevant yeasts. With the exception of a few species (e.g. C. glabrata for azoles and C. parapsilosis for echinocandins), the findings of our study suggest that two of the most widely used commercial methods (Etest and Sensititre) provide valid and reproducible results.
BMC Infectious Diseases 01/2011; 11:130. · 3.12 Impact Factor
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ABSTRACT: We conducted a retrospective survey of nocardiosis in 9 city hospitals in northern Italy from 1982 to 1992. The medical records of 30 patients with documented nocardiosis were reviewed. Microbiological data included morphology, biochemical characteristics, serology and in vitro susceptibility testing. The 29 isolates (1 case was diagnosed on the basis of serological results) were Nocardia asteroides (n = 25) and Nocardia farcinica (n = 4). Predisposing factors included immunosuppression for organ transplant rejection prophylaxis, lung disease (silicotuberculosis and pulmonary fibrosis), solid tumours and hematological malignancies, and AIDS. Three patients had no identified risk factors. 20 cases of pulmonary nocardiosis were observed. Sites of infection in patients without previous pulmonary involvement were: brain abscesses, soft tissues, pericardium, blood, and cerebrospinal fluid. Most strains tested were susceptible to amikacin and imipenem. Resistance to several antimicrobial agents was found, particularly erythromycin, fosfomycin, pefloxacin, sulphonamides and trimethoprim. Antimicrobial chemotherapy included sulphonamides, amikacin, ceftriaxone, imipenem and minocycline. 21 patients survived, although 2 relapsed transiently. Nocardiosis appears to be more common than generally realised by physicians in northern Italy. The local species distribution and disease spectrum are similar to those described elsewhere. Nocardiosis should be part of the differential diagnosis in patients with pulmonary infiltrates or brain abscess, particularly those with predisposing factors.
07/2009; 27(1):23-27.
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ABSTRACT: Cladophialophora boppii is a dematiaceous fungus, which has been reported only rarely to be the cause of cutaneous infection. Herein we describe a C boppii parenchymal and bronchial infection in a lung transplant recipient. We also illustrate the clinicoradiologic patterns and review possible treatment options for these difficult infections.
The Journal of heart and lung transplantation: the official publication of the International Society for Heart Transplantation 07/2009; 28(6):635-7. · 3.54 Impact Factor
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ABSTRACT: Acinetobacter baumannii is typically a nosocomial pathogen. Epidemiologic tools that can rapidly trace the spread of hospital-associated infections due to this microorganism are essential. Currently, amplified fragment length polymorphism and pulsed-field gel electrophoresis using ApaI, a macrorestriction enzyme, are the molecular techniques most widely used to type this microorganism. Unfortunately, they are technically demanding, requiring also well-trained personnel, and are time consuming. The aims of this study are 1) to evaluate the usefulness of the semiautomated repetitive-sequence-based polymerase chain reaction (rep-PCR) for typing A. baumannii, comparing this method with another semiautomated technique, such as ribotyping, and 2) to acquire information about the incidence, the clinical significance, and the susceptibility patterns of this microorganism in 13 different Italian hospitals in a 4-week period (total study population, >14000 beds). Twenty-eight A. baumannii were isolated in 7 different hospitals; 21 strains were analyzed with molecular methods. Automated ribotyping distinguished 6 different clusters of isolates, whereas rep-PCR appeared to be more discriminating, allowing us to distinguish 8 different clusters. Our study confirms the good discriminatory power of the semiautomated rep-PCR. Although expensive, this method is simple, fast, and reproducible, and in our opinion, it could be used in a hierarchic approach as a 1st-line typing tool if results of analysis are required in a short period or if a large number of isolates have to be analyzed.
Diagnostic Microbiology and Infectious Disease 01/2008; 60(1):1-7. · 2.53 Impact Factor
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ABSTRACT: Paranasal sinus fungus ball is an extramucosal mycosis, usually occurring in immunocompetent people as a monolateral lesion. To review the literature data and to report the Policlinico S. Matteo, University of Pavia experience, 81 patients presenting paranasal fungus ball have been treated (January 1994 to May 2005). Twenty-seven men and 54 women (19-91 years old; mean 49.4 years) were considered. Seventy-three patients had a single sinus affected, but eight presented multiple localisations. Maxillary was the most involved sinus followed by sphenoidal and ethmoidal. Moulds have been isolated in 28/81 cases. Histology showed fungal colonisation but not invasion in all cases. Tomography showed bone erosion in 33.3% of patients. All have been treated only by functional endoscopic sinus surgery. Seventy-seven of 81 patients have been cured. Four of 81 patients needed another surgical treatment. Follow up was between 6 and 132 months (average: 63 months). Fungus ball is a sinusal pathology caused by mycetes like Aspergillus spp. Histology confirms the fungal aethiology excluding tissue invasion. Mycological culture consented to identify the pathogenic mould in 34.5% of cases. Actually functional endoscopic sinus surgery is the gold standard for treatment of this pathology, and antifungal therapy is unnecessary.
Mycoses 12/2007; 50(6):451-6. · 2.25 Impact Factor
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ABSTRACT: The purpose of this study was to verify the standard procedures and minimum level of knowledge of Italian public laboratories involved in the management of antifungal susceptibility testing (AST). Two nationwide surveys were performed in 1999 and 2004. One hundred and two Italian hospitals located in 85 provincial capitals (82.5%) participated to these surveys. In 1999, 28 (27.5%) laboratories versus 16 (15.7%) in 2004 stated that they did not perform any susceptibility testing. Some discrepancies observed in the survey confirm that AST is difficult to be correctly managed, and that it can be performed only in very well-trained centers. The great variability of the results of MIC determination and clinical interpretation underlines the urgent need to improve knowledge about indications, method choice, and interpretative criteria for AST both for clinical microbiologists and clinicians.
Diagnostic Microbiology and Infectious Disease 03/2007; 57(2):225-7. · 2.53 Impact Factor
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ABSTRACT: In the past, no case reports concerning N. brasiliensis infections were published from Italy. We now report 4 cases observed during 1998-2006 in 4 Italian patients, 1 immunosuppressed and 3 immunocompetent.
Scandinavian Journal of Infectious Diseases 02/2007; 39(11-12):969-74. · 1.72 Impact Factor
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ABSTRACT: In the past the Italian soil was considered as a low-endemic pabulum for H. capsulatum var. capsulatum and only few autochthonous cases of histoplasmosis were reported in Italy, especially in the Po valley. The aim of the paper was to evaluate this possibility by reviewing the literature and providing our own personal data. Four additional cases of histoplasmosis were observed during 1999-2003 in AIDS immigrant or in Italian citizens, and in travellers to endemic areas. One of the AIDS patients was an autochthonous case of histoplasmosis. The Italian literature was reviewed. Recent cases and literature data confirm the possible autochthonous presence of histoplasmosis in Italy, especially in the Northern regions.
Revista Iberoamericana de Micología 10/2005; 22(3):169-71. · 1.16 Impact Factor
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ABSTRACT: The aim of this study was to evaluate the acridine orange leukocyte cytospin (AOLC) test for the rapid diagnosis of septicemia caused by central venous catheters (CVCs), without removing the catheter, in a pediatric intensive care unit population. Twenty-six patients admitted in the pediatric intensive care unit of Azienda Ospedaliera "Ospedali Riuniti di Bergamo", Italy, were prospectively evaluated for CVC-related infection. Blood for culture was taken from all patients. Quantitative endoluminal cultures of the removed catheter tip by Cleri's technique and semiquantitative superficial cultures of the hub were performed. Gram staining and an AOLC smear were done according to Kite's technique. Four Staphylococcus CVC-related bloodstream infections were identified. CVC colonization was detected in 8 patients. Four had septicemia (Enterococcus faecalis, Escherichia coli, Klebsiella oxytoca, Candida glabrata) without CVC involvement. However, Gram staining and the AOLC test were negative in all cases. We conclude that cytocentrifugation and acridine orange staining of blood withdrawn by Kite's method from an in situ catheter, although simple, quick, and inexpensive, did not aid diagnosis in this pediatric population.
Diagnostic Microbiology and Infectious Disease 09/2005; 52(4):337-9. · 2.53 Impact Factor
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ABSTRACT: We performed an open study to investigate the use of electrical stimulation (ES) on the vestibular area and vaginal introitus in women with sexual pain disorders. We recruited 29 women (age range 20-45 years) from among the patients at our Reproductive Psychobiology Unit to participate in the present study. They each experienced vestibular pain, inducing dyspareunia and vaginism. We performed ES with an ECL43400 apparatus (Elite, EssediEsse srl, Milan, Italy) once a week for 10 weeks. To evaluate the muscular activity of the perineal floor and sexual function, we employed the same apparatus with a vaginal probe for recording myoelectrical activity (muV), we employed a VAS scale for evaluating pain, and we administered the Female Sexual Function Index (FSFI; Rosen et al., 2000) before and after the study protocol. We analyzed data by parametric and nonparametric comparisons and correlations, as appropriate. Our major findings were as follows: (a) the contractile ability of pelvic floor muscles (p < 0.001), as well as the resting ability (p < 0.001), significantly improved following ES; (b) the current intensity tolerated significantly increased (p < 0.001) throughout the study, from 41.3 +/- 7.4 mA at the start of the study to 50 +/- 7.4 mA at the end of the stimulation protocol; (c) the Visual Analogic Scale (VAS) for pain significantly declined (p < 0.001), whereas FSFI pain scores (p < 0.001) and full scale scores (p < 0.001) significantly improved following ES, and 4 out of 9 women with vaginism went back to coital activity; (d) FSFI pain score and the current intensity tolerated, both before (R = .59; p < 0.006) and at the end (R = .53; p < 0.02) of the stimulation protocol, positively correlated. ES may be effective in the management of sexual pain disorders. Further controlled studies are necessary to standardize stimulation protocols according to the severity of pain and to better clarify the long-term clinical effects of ES.
Journal of Sex and Marital Therapy 01/2003; 29 Suppl 1:103-10. · 1.27 Impact Factor
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ABSTRACT: Scedosporium apiospermum is an environmental mould. Human infection caused by this organism is described more and more often. However, only a few case reports demonstrate its role as a telluric contaminant in patients affected by traumatism. This report is the case of a severe post traumatic infection by S. apiospermum in an immunocompetent young man. Surgical drainage associated with systemic therapy was successful.
Brain Injury 08/2002; 16(7):627-31. · 1.36 Impact Factor
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ABSTRACT: Invasive aspergillosis is a serious problem for immunocompromised patients, especially if neutropenic. The diagnosis of this infection is complicated, since clinical symptoms are often similar to those of other fungal diseases. The chance of detecting the presence of a specific antigen in the serum could confirm the suspected clinical diagnosis and. perhaps, be useful for the follow-up of the patient. The Medical Mycology Committee of the Associazione Microbiologi Clinici Italiani (AMCLI) decided to evaluate in a multicenter prospective study (from I November 1998 to 28 February 1999) the performance of the Platelia Aspergillus Kit (Bio-Rad) for the detection of Aspergillus galactomannan in human serum. The enrolled patients included various groups of immunosuppressed patients (mostly neutropenic). Blood samples were drawn at the time of enrollment. This decision was based upon a clinical diagnosis of probable aspergillosis (antibiotic non-responsive fever for at least 96 hours, cough, hemophthosis and positive chest X-ray). Additional blood samples were drawn on days 3, 6, 9, 12, 15 and 21. Culture and histopathologic examinations were performed according to the individual laboratory workflow. For each patient the laboratory filled a form with all the available clinical information, to create a database on which to evaluate the results of the test. During the study, 187 patients with various kinds of immunosuppression were enrolled. A total of 256 sera were tested: for 117 patients (62.6%) only the basal sample was tested, whereas for the 70 symptomatic patients (37.4%) multiple specimens (range: 1-6) were tested. The results allowed the laboratories to exclude (68.6%) or confirm (31.5%: confirmed and/or probable) the clinical diagnosis of invasive aspergillosis; 4 cases remained undetermined. Based on the results of this study, it seems that the use of this test should be limited to those patients with clinical symptoms of aspergillosis.
Mycopathologia 02/2002; 155(3):129-33. · 1.65 Impact Factor
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Clinical Microbiology and Infection 07/1996; 1(4):276-278. · 4.54 Impact Factor
