HB&L - Alfred 60 Scientific Pubblications

MICROBIOLOGIA MEDICA, Vol. 25 (3), 2010

Evaluation of an automated method for urineculture screening

Claudia Ballabio1, Natascia Venturi1, Maria Roberta Sala1, Paolo Mocarelli1, Paolo Brambilla1, 2

1 Servizio Universitario di Medicina di Laboratorio Ospedale di Desio, Desio
2 DIMS, Facoltà di medicina e Chirurgia, Università Di Milano, Bicocca

SUMMARY
Introduction: Urinary tract infections are one of the most common diseases found in medical practice and are diagnosed with traditional methods of cultivation on plates. In this study we evaluated an automated instrumentation for screening of the urineculture that can provide results quickly and guarantee traceability. The comparison of results obtained with automatic and plate methods is reported.
Methods: 316 urine samples including midstream urine, urine catheter and urine bag have been analyzed by Alfred 60 (Alifax) through light scattering technology that measures the replication of the bacteria. Simultaneously, the samples were sown on agar plates CPS3,Agar Cled, Mc Conkey Agar.
Results: A total of 316 samples were analyzed by the automated method, 190 resulted negative, all confirmed by culture, while 126 were found positive. 82 cases were confirmed positive in culture plate, 65 with significant isolation of bacteria and 17 with poly-microbial flora with a significant charge. 44 cases were negative in culture plate but positive for the automated method.
Conclusions: The absence of false negative results at low charges can represent a starting point to introduce an automated method for urinocolture screening.

Key words: Urinary tract infections,Automatization, Urineculture

MICROBIOLOGIA MEDICA, Vol. 25 (2), 2010

Evaluation of the HB&L system for the culture of prosthetic and osteoarticular origin samples

Agostina Ronca1, Sabrina Brenci1, Giuliana Carrega2, Gianni Riccio2, Luisa Santoriello1

1 Unità Dipartimentale Semplice di Microbiologia-ospedale Santa Corona - Asl 2-Pietra ligure (SV)
2 Unità Complessa di Malattie Infettive-Ospedale Santa Corona - Asl2 Pietra Ligure (SV)

SUMMARY
Prosthetic and osteoarticular infections represent a complex condition to diagnose and resolve. In both cases, the eradication of microorganisms is difficult because of anatomical and physiological characteristics of the site of infection (bone). The best strategy for an effective pharmacological treatment is based on an early diagnosis confirmed by microbiological testing of bone, periprosthetic tissue or removed prostheses, to support clinicians to undertake prolonged targeted therapy. The purpose of this paper is to assess the clinical correlation between the results of the cultures performed with automated HB&L (ALIFAX) system compared to the result of traditional methods. HB&L is a system to perform bacterial cultures, susceptibility and direct RAA tests on biological materials based on detection of bacteria and fungi by laser light-scattering kinetics in liquid culture medium at 37°C. Samples get to laboratory are inoculated in rich broth and placed in the incubator. The next day are processed according to the following protocol: 500 ml of the broth are transferred in the vial of the instrument and 200 ml of supplement DEB for fastidious organisms are added. The cultures are incubated for 360 minutes in the same time PAR test (antimicrobial residual power) is determined. In the first half of 2009 418 samples collected from prostheses and osteoarticular infections from 118 patients were analyzed. The tests showed 304 negative and 114 positive samples (27.30%) from which were isolated Gram negative (17.5%) and Gram positive (82.5%) bacteria. Gram positive included 38.3% S. aureus, 33% coagulase negative Staphylococci and 26.6% Enterococci. Preliminary data obtained by the HB&L system for the culture of tissue sample, as well as the improvement of surgical techniques have led to a significant increase in correlation with the clinical data compared to traditional microbiological analysis.

MICROBIOLOGIA MEDICA, Vol. 25 (1), 2010

Preliminary indications for antibiotic susceptibility tests in less than six hours in positive blood cultures

Vesselina Kroumova, Elisa Gobbato, Paola Macaluso, Sabrina Tamburelli, Francesca Marini, Marcella Perone, Stefania Orlandi, Morena Viviani, Giacomo Fortina

Laboratorio di Microbiologia e Virologia Azienda Ospedaliera Universitaria "Maggiore della Carità”,

SUMMARY
A rapid determination of the antibiotic susceptibility patterns of pathogens responsible for sepsis represents a significant milestone for a timely correct antibiotic therapy. The system HB&L® (ALIFAX) allows reduced time in the detection of bacterial growth and consequently is able to detect the growth or absence of certain microorganisms in the presence of a given antibiotic. In this study three system for rapid antibiotic susceptibility tests among bacteria isolated from blood were compared: HB&L® (ALIFAX),VITEK®2, (BioMérieux) and essays Etest® (BioMérieux). Present findings indicate that HB&L® (ALIFAX) is rapid reliable instrument that may support the clinician for a rapid and appropriate treatment, particularly in the critical patient.

Key words: Antimicrobial susceptibility tests; Sepsis, Critical illness.

MICROBIOLOGIA MEDICA, Vol. 25 (1), 2010

HB&L System: rapid determination of antibiotic sensitivity of bacteria isolated from blood cultures

Simone Barocci, Marinella Giacomini, Antonella Renzi, Marcello Palma, Luciano Latini, Luciano Quagliarini, Antonio Migali

ASUR MARCHE z.t. 4 Senigallia, U.O. Patologia Clinica

SUMMARY
Introduction. Blood culture is an important method to detect microbial pathogens on blood, very useful for diagnosing bacterial infections. Unfortunately, classical diagnostic protocols cannot directly identify bacteria responsible for sepsis and accordingly their antimicrobial profiles. This problem causes a delay of almost two days in the availability of a specific antimicrobial profile. Objective. Among the main causes of death, sepsis have a relevant importance. For this reason it is important both to identify pathogens and to perform an antimicrobial susceptibility test in the shortest time as possible. For this purpose, the main aim of this study is the evaluation of the performances of an antimicrobial susceptibility determination directly performed on positive blood cultures. Materials and methods. This study has been performed on 70 positive blood cultures, during the period from January to July 2009. A number of 35 blood cultures were positive for Gram negative bacteria, and 35 were positive for Gram positive bacteria. From these positive blood cultures, after a short sample preparation, it has been possible to directly determine antimicrobial susceptibility profiles by using the HB&L (formerly URO-QUICK) instrument. Results. The HB&L system results showed a very good correlation with both the classical disk diffusion method and VITEK 2 automatic system. The performances between the methods carried out in this study were equivalent. Conclusions. From data reported, thanks to the rapidity and simplicity of the method used, we can assert that the direct susceptibility test available with the HB&L system, is useful for a rapid and early choice of the antibiotic treatment.

Key words: Antimicrobial Susceptibility Test, blood culture, EUCAST.

New Microbiol. 2010 Apr;33(2):147-53.

Rapid reporting of urine culture results: impact of the uro-quick screening system.

Ilki A, Bekdemir P, Ulger N, Soyletir G.

Marmara University, Medical Faculty, Department of Microbiology, Haydarpasa, Istanbul, Turkey. ailki@superonline.com

ABSTRACT
This study evaluated the impact of the Uro-Quick (UQ) screening system (Alifax, Italy) for a rapid and accurate reporting of urine cultures, and whether it can provide bacterial yield to be used in identification and susceptibility testing. A total of 1480 urine samples collected between October 2006 and July 2008 were tested by conventional culture (CC) methods and UQ simultaneously. Sediments of positive UQ vials were used as bacterial yields for identification and susceptibility testing procedures. Of the 1480 samples, 999 revealed bacteria and/or leukocytes in direct microscopy. Among these 999 samples, positive growth was detected in 420 (42%) and 433 (43.3%) by UQ and CC, respectively. However, only 0.6% of samples without bacteria and leukocytes exhibited positive growth. When compared to CC, UQ demonstrated high levels of positive predictive value (95.9%), negative predictive value (94.8%), sensitivity (93%) and specificity (96.9%). Both CC isolates and UQ bacteria showed 81.3% concordance in identification results. Susceptibility testing of UQ bacteria displayed >90% agreement, when compared with standardized disk diffusion test. Our results indicate that UQ can reliably be used in routine laboratories giving microbial growth results in 3 hours. The most significant part of the study is that bacterial yields of UQ positive samples can be used in identification and susceptibility testing, allowing a rapid, same-day reporting of urine cultures.

PMID: 20518276 [PubMed - indexed for MEDLINE]

J Microbiol Methods. 2010 Jun;81(3):235-9. Epub 2010 Mar 27.

Evaluation of the Uro4 HB&L system for the rapid diagnosis of lower respiratory tract infections in intensive care units

Andrea T., Laura S., Antonietta C., Giuseppe PS., Mario C., Giorgio P.

Department of Histology, Microbiology and Medical Biotechnologies, University of Padova, Center of Community Medicine, ULSS20 Verona, Italy.

ABSTRACT
Respiratory tract infection is a common and important problem in the intensive care unit (ICU) setting. It has been demonstrated that an appropriate initial regimen or its early modification (within 6-12h from diagnosis) based on microbiological results leads to a higher survival rate. Here we evaluated the Uro4 HB&L automated system for the rapid diagnosis of respiratory tract infections in ICU patients. A total of 644 lower respiratory tract specimens collected from 400 inpatients from nine ICUs at the Padova University hospital were collected during a 12-month period. All samples were processed both with the Uro4 HB&L system and with the reference culture method. Out of 322 samples, 312 were concordant positive, 276 out of 276 were concordant negative, 66 samples were declared uncertain and discarded because of an excess in turbidity. The diagnostic accuracy was good, compared with standard cultures from BAL specimens, in terms of sensitivity (0.972), specificity (1.00), likelihood ratios and diagnostic odds ratio. Ten discordant samples, resulted positive with the reference culture and not detectable with the Uro4 HB&L, were confirmed positive by Gram-stain smear analysis performed after incubation. The Uro4 HB&L system, compared to the standard culture method, revealed a very high sensitivity and a full specificity in identifying clinically relevant microorganisms from lower respiratory tract samples after merely 6h. Overall our results indicate that Uro4 HB&L is a reliable system for the surveillance of the respiratory tract infections in ICUs; it could speed up the laboratory procedures and provide fast, reliable results for clinicians.

2010 Elsevier B.V. All rights reserved.
PMID: 20347888 [PubMed - indexed for MEDLINE]

Oral Presentation, 20th ECCMID, Vienna, 10-13 April 2010.

Speeding up identification of bacteria from urine by the use of an Alifax HB&L Uroquattro incubator followed by Maldi Biotyper identification

M. Kostrzewa, C. Boogen, J. Maier, U. Weller (Bremen and Cologne University, Germany)

Objectives: Direct identification of bacteria from infected urine by the MALDI-Biotyper (MBT) has been shown previously. To enable quantification as well as identification, which is necessary for routine diagnostic we have tested the combination of an automated quantification system, the HB&L Uroquattro (ALIFAX HBL), with the MALDI-TOF workflow.

Methods: For this proof of concept study, 161 urine samples from routine were analysed in parallel. For the novel combined workflow, a 500 μl aliquot was transferred into a 2ml HB&L tube with nutrient broth and stirrer, placed into the HB&L instrument for 3 h at 37°C and the CFU value was read. Turbidity of the vials is monitored and bacterial count is calculated. After incubation, 500 μl aliquots from the positive vials were pipetted onto a 100 μl Ficoll-Paque Plus cushion and centrifuged (5 min at 15000 g in a 1.5 ml Eppendorf cup). Supernatant was discarded and part of the pellet transferred onto a MALDI target, air dried, overlayed with matrix solution (HCCA), again air dried and subjected to identification by mass spectrometry using the MBT system. This approach was compared to our standard workflow: plating the urine on 2 Petri dishes in a quantitative way, MBT identification of colonies after over night incubation at 37°C.

Results: A bacterial count above 10E4 CFU was obtained for 77 of the 161 samples. These patients where considered to have a bacterial infection. The bacterial count was identical to a large extend with both methods. However, MBT identification was only possible in 63 of the samples incubated in the HB&L. Here, the results matched those of the standard protocol. This was due to the presence of more than one pathogen (as shown by the standard workup) which the current identification software of the MBT can not resolve.

Conclusion: This protocol enables reliable bacterial identification together with a bacterial count in less than 3 hours in cases of mono-bacterial infection using the standard protocol of the HB&L. Ongoing improvement to the MBT identification algorithms will resolve the problem of poly-bacterial infections. Further extension of this approach to usually sterile body fluids, tracheal secretions and pleural aspirates also here may lead to a dramatic reduction in turnaround time of microbiological labs for identification and subsequent AST-testing.

Med Sci Monit. 2009 Feb;15(2):BR55-60.

A novel culturing system for fluid samples.

Fontana C, Favaro M, Minelli S, Bossa MC, Altieri A, Favalli C.

Department of Experimental Medicine and Biochemical Sciences, Tor Vergata, University of Rome, Rome, Italy. carla.fontana@uniroma2.it

Abstract
BACKGROUND: Large-volume culture methods for sterile body fluids employing automated blood-culture systems increase the recovery of microorganisms compared with traditional plate medium methods. However, in many instances a laboratory receives only small-volume samples.

MATERIAL/METHODS: The URO-QUICK system (now HBandL), originally used to process urine samples, was evaluated for organism enrichment and determination of microbial count of fluid samples. Fluid specimens were also evaluated for their residual antimicrobial activity (RAA). The procedures were compared with results from a conventional culture procedure. The 546 samples included 106 endotracheal aspirate, 63 bronchoalveolar lavage, 139 sputum, 47 blood, 105 pleural fluid, 26 cerebrospinal fluids, and 41 peritoneal fluid samples as well as 19 other fluids including synovial fluid (n=5), ascitic fluid (n=9), fluids from the drainage of an infected central venous catheter (n=3), abdominal drainage fluid (n=1), and cholecystic fluid (n=1).

RESULTS: The URO-QUICK system allowed the culture of an additional 44 samples (8%, p=0.007) compared with the traditional culturing method. The RAA test demonstrated good concordance with the reference method, showing specificity and positive predictive value of 100% for each, while the sensitivity and the negative predictive value were 67% and 76%, respectively. The microbial counts evaluated using the URO-QUICK system showed excellent agreement with traditional enumeration methods.

CONCLUSIONS: The URO-QUICK system may well represent an excellent alternative to solid medium-based recovery and enumeration methods.

PMID: 19179962 [PubMed - indexed for MEDLINE]

Annals of Microbiology, 56 (2) 179-183 (2006)

Evaluation of the Uro-Quick system for antibiotic susceptibility tests of strains collected from intensive care units

Elisabetta PEZZATI, Sonia MARENGO, Simona ROVETA, Clara CASSANELLI, Elisabetta MAIOLI, Fabrizio CAVALLINI, Simone CAGNACCI, Laura GUALCO, Anna MARCHESE, Eugenio A. DEBBIA*

Institute of Microbiology, DISCAT, University of Genoa, Largo Rosanna Benzi 10, 16132 Genoa, Italy

Abstract
During the period January-June 2004, 525 pathogens isolated from intensive care units were examined with the new rapid Uro-Quick method for antibiotic susceptibility tests. The results were compared with those obtained by the reference NCCLS methods (disk diffusion or dilution). Antibiotic (in appropriate concentration) was introduced in a vial containing 2 ml of Mueller-Hinton broth, then 0.5 ml of 5 x 105 or 106 cells/ml of the strain culture were added. After 3-6 h of incubation, depending on the microorganism studied, the instrument printed the results: no growth and a growth curve similar to that of the untreated control are representative of a susceptible and resistant strain respectively. The following drugs were tested: ciprofloxacin, ampicillin, aztreonam, co-clavulanate, piperacillin/tazobactam, ceftazidime, cefotaxime, cefuroxime, ceftriaxone, imipenem, amikacin, gentamicin, trimethoprim-sulfamethoxazole, clindamycin, erythromycin, linezolid, penicillin, tetracycline, vancomycin, oxacillin. Gram-negative strains tested were 252 and Gram-positive 273: agreement between the two methods ranged from 85.6% (piperacillin/tazobactam) to 98.5% (ciprofloxacin) in Gram-negative pathogens, from 90 to 100% in Gram-positive, with the exception of erythromycin (84.2%) against enterococci. On the basis of the present findings the Uro-Quick system appears to be very useful for the rapid detection of antibiotic susceptibility in pathogens collected from intensive care units.

Key words: Uro-Quick, antibiotic susceptibility tests, disk-diffusion method, intensive care units.

J Chemother. 2004 Apr;16(2):107-18.

Evaluation of the Uro-Quick, a new rapid automated system, for the detection of well-characterized antibiotic-resistant bacteria.

Roveta S, Marchese A, Debbia EA

Sezione di Microbiologia-DESCAT. Università degli studi di Genova. Largo Rosanna Benzi 10-16132, Genoa, Italy

Abstract
The Uro-Quick system has been employed to detect antibiotic resistance in genotypically and/or phenotypically well-characterized bacterial species including those that might not be easily identified by routine procedure. In order to achieve full agreement between the antibiotic susceptibility results obtained by the reference method (NCCLS) and the Uro-Quick system, the optimal experimental conditions (inoculum size, time of incubation and antibiotic concentration) for each strain to be used by the automatic system were determined. The shorter time periods for generation of correct susceptibility results were 180 min for ampicillin- and ciprofloxacin-resistant Escherichia coli and for ESBL- and Inhibitor-resistant TEM (IRT)-producing E. coli; 360 min for penicillin-susceptible Streptococcus pneumoniae, as well as for strains with reduced susceptibility to this antibiotic (both intermediate, and resistant isolates). The same time was required to detect erythromycin-resistant pneumococci irrespective of their mechanism of resistance (ribosomal methylation and efflux-mediated), Streptococcus pyogenes exhibiting the three erythromycin-resistance phenotypes (constitutive, inducible and M-type) and Klebsiella pneumoniae, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Proteus mirabilis and Moraxella morganii refractory to third-generation cephalosporins, aminoglycosides, ciprofloxacin and other classes of antimicrobial agents; 480 min for penicillin-resistant, constitutive and inducible oxacillin-resistant (OXA-R) Staphylococcus aureus and OXA-R Staphylococcus epidermidis. The same period of time was also necessary to find the great majority of drug-resistance exhibited by Pseudomonas aeruginosa. Teicoplanin-resistant Staphylococcus haemolyticus, vancomycin-resistant (VanA, VanB, VanC) high-level aminoglycoside-resistant (HLAR) Enterococcus spp, and imipenem-resistant P. aeruginosa required longer incubation (24 h) to be detected. The results obtained indicate that Uro-Quick might be a reliable and promising instrument for the correct detection of the above antibiotic resistance markers.

PMID: 15216942 [PubMed - indexed for MEDLINE]

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