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Giavarina D.,San Bortolo Hospital | Villani A.,Clinical Laboratory | Caputo M.,Clinical Chemistry and Microbiology Laboratory
Biochemia Medica | Year: 2010

Part of this document has been endorsed as a Position Statement on Point of Caretesting (in-hospital setting) of the Italian Society of Laboratory Medicine (Società Italiana di Medicina di Laboratorio, SIMeL) and also refers to offcial documents and International standards to for generalities (ISO 15189/2003) and specific items (ISO 22870/2006). As such, this article is ba sed on to professional standards, guidelines and peer reviews documents, and it is aimed to im prove the pre-analytical, analytical and post-analytical phase of point of caretesting (POCT), by providing insights into definitions, key aspects in developing a diagnostic system for POCT, benefits and risks of POCT and leading sources of errors. Source

Torretta S.,University of Milan | Marchisio P.,University of Milan | Colombo M.R.,Clinical Chemistry and Microbiology Laboratory | Rosazza C.,University of Milan | Pignataro L.,University of Milan
European Journal of Clinical Microbiology and Infectious Diseases | Year: 2016

The purpose of this investigation was to describe the first application of nasopharyngeal cytology (NPC), a new cytological technique to collect cellular material from the nasopharyngeal surface non-invasively in children with chronic adenoiditis associated with recurrent acute otitis media and/or otitis media with effusion. Cellular material was collected transorally using an extra-thin flexible wire nasopharyngeal swab and then examined under a light microscope. The diagnostic accuracy of NPC in detecting the presence of allergy and pathogens (compared to microbiological evaluation of nasopharyngeal aspirates, NPAs) was assessed. NPC was performed on 121 children (mean age 69.4 ± 15.5 months). Inflammatory cells and pathogens were detected in 61.1 % and 44.2 % of patients, respectively. The specificity of nasopharyngeal eosinophils in detecting allergy was good (91.9 %), as was the specificity of mast cells, but the sensitivities were less. The NPAs revealed bacterial colonisation in 84.7 % of the patients, and Streptococcus pneumoniae was the most frequently isolated (60.0 %). NPC revealed the presence of bacteria in 94.9 % of patients, including bacillary species in 33.9 %. NPC was highly sensitive in detecting pathogens (96.0 %). Its specificity in detecting bacillary species was fairly good (75.0 %), but the corresponding values of the specificity of NPC in detecting pathogens and its sensitivity in detecting bacillary species were poor. Our findings suggest the need for more structured studies that can test the real effectiveness and usefulness of NPC in defining nasopharyngeal cytological patterns in children with chronic nasopharyngeal diseases by comparing it with established diagnostic techniques. © 2016, Springer-Verlag Berlin Heidelberg. Source

Gessoni G.,Madonna della Navicella Hospital | Saccani G.,Clinical Chemistry and Microbiology Laboratory | Valverde S.,Madonna della Navicella Hospital | Manoni F.,Civil Hospital | Caputo M.,Clinical Chemistry and Microbiology Laboratory
Clinica Chimica Acta | Year: 2015

Urine culture is the most frequently requested test for a Microbiology Lab. A reliable screening tool would be of paramount importance both to clinicians and laboratorians, provided that it could get fast and accurate negative results in order to rule-out urinary tract infection (UTI). Materials and methods: We evaluated 1907 consecutive urine samples from outpatients. Culture was performed on chromogenic agar with 1μL loop, using 105CFU/mL as a limit of positive growth. Using Sysmex Uf-1000i analyzer we evaluated bacteria forward scatter (B_FSC) and fluorescent light scatter (B_FLH) in a preliminary discrimination step for UTI caused by Gram+ or Gram- bacteria. Results: We got 512 positive samples. A mono-microbial infection was observed in 490 samples; two bacterial strains were isolated in 22 samples, so 534 bacterial strains were found: 392 Gram-, 133 Gram. + and 9 yeasts. Comparing Gram. + and Gram. - bacteria we observed a statistically significant difference for B_FSC but not for B_FLH. In this application experimental cut-off value for B_FSC was 25ch. Using this cut-off to perform a presumptive identification of UTI sustained by Gram-+ bacteria, we observed a SE 0.68, SP 0.84. Conclusion: Our data although preliminary suggest that B_FSC could be useful in presumptive exclusion of UTI caused by Gram-positive bacteria. © 2014 Elsevier B.V. Source

Mastronicola D.,CNR Institute of Molecular Biology and Pathology | Testa F.,CNR Institute of Molecular Biology and Pathology | Forte E.,CNR Institute of Molecular Biology and Pathology | Bordi E.,Clinical Chemistry and Microbiology Laboratory | And 3 more authors.
Biochemical and Biophysical Research Communications | Year: 2010

Flavohemoglobins (flavoHbs), commonly found in bacteria and fungi, afford protection from nitrosative stress by degrading nitric oxide (NO) to nitrate. Giardia intestinalis, a microaerophilic parasite causing one of the most common intestinal human infectious diseases worldwide, is the only pathogenic protozoon as yet identified coding for a flavoHb. By NO amperometry we show that, in the presence of NADH, the recombinant Giardia flavoHb metabolizes NO with high efficacy under aerobic conditions (TN=116±10s-1 at 1μM NO, T=37°C). The activity is [O2]-dependent and characterized by an apparent KM,O2=22±7μM. Immunoblotting analysis shows that the protein is expressed at low levels in the vegetative trophozoites of Giardia; accordingly, these cells aerobically metabolize NO with low efficacy. Interestingly, in response to nitrosative stress (24-h incubation with ≥5mM nitrite) flavoHb expression is enhanced and the trophozoites thereby become able to metabolize NO efficiently, the activity being sensitive to both cyanide and carbon monoxide. The NO-donors S-nitrosoglutathione (GSNO) and DETA-NONOate mimicked the effect of nitrite on flavoHb expression. We propose that physiologically flavoHb contributes to NO detoxification in G. intestinalis. © 2010 Elsevier Inc. Source

Saccani G.,Clinical Chemistry and Microbiology Laboratory | Gessoni G.,Madonna della Navicella Hospital | Valverde S.,Madonna della Navicella Hospital | Manoni F.,Civil Hospital | Visconti M.,Clinical Chemistry and Microbiology Laboratory
Biochimica Clinica | Year: 2014

Urine culture is the most frequent test in microbiology laboratories. A screening tool, providing fast and reliable results to rule-out urinary tract infection (UTI), would be of great importance. We studied 1043 consecutive urine samples by Sysmex UF-1000i analyzer. Comparison was made by robotic urine culture on chromogenic agar with 1 μL loop, using 105 CFU/mL as a limit of positive growth. We evaluated bacteria quantification for rapid exclusion of UTI and bacteria forward scatter (B-FSC) in preliminary discrimination of UTI caused by Gram positive or Gram negative bacteria. For exclusion of UTI, the best cut-off value was 130 bacteria/μL. At this threshold, the sensitivity (SE) was 0.98 and the specificity (SP) 0.75. For exclusion of UTI sustained by Gram positive bacteria, the best cut-off value for B-FSC was 25ch. At this threshold, SE was 0.68 and SP was 0.89. Source

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