Daria Pašalić
Department of Medical Chemistry, Biochemistry and Clinical Chemistry
Zagreb University School of Medicine
Šalata ul 2.
10 000 Zagreb, Croatia
Phone +385 (1) 4590 205; +385 (1) 4566 940
E-mail: dariapasalic [at] gmail [dot] com

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Screening for the urinary tract infections using Sysmex UF 1000i flow cytometer
Konderak J (1), Elmardi A (2), Farkas B (2), Pinter E (1)
(1) Synlab Hungary Ltd., Medical Biochemical Department, Budapest, Hungary
(2) Synlab Hungary Ltd., Microbiological Department, Budapest, Hungary
Corresponding author: jkonderak [at] gmail [dot] com
Background: Bacterial cultures for the urinary tract infections (UTI) are the most common microbiological tests. Huge amounts of negative cultures demand for effective screening method reducing cost and time. The aim of this study was to evaluate the efficacy of UF 1000i flow cytometer for preselecting of negative results.
Material and methods: Urine specimens (N = 1226) suspected UTI were simultaneously cultured and analyzed by UF 1000i for bacterial (BC) and leukocyte (WBC) counts. Population: male:female = 1:2; ages: 0-15 years: 8%; 16-65 years: 59%; > 65 years: 33%; outpatient:inpatient = 4:1. For culture samples were inoculated using a 10 μL loop on selective agar plates. After standard incubation results were evaluated. A sample was considered negative for UTI if growth was < 103 CFU/mL (colony forming unit). Positive samples were attributed to one of the levels of CFU/mL (> 103, > 104, > 105).
Results: Using culture results as gold standard, we performed ROC analysis to determine area under curve (AUC), cutoff values (CO), sensitivities (SE), specificities (SP), negative predictive values (NPV) in pointing to BC and WBC measured by UF1000i. Results of ROC statistics at CFU/ml of > 103, > 104, > 105 AUC: 0.88, 0.91 and 0.93, CO BC and/or WBC/µL: > 9, > 22, > 100, SE%: 95, 96, 96; SP%: 31, 45, 54; NPV%: 97, 98, 98, respectively.
Conclusions: By screening out method, UF 1000i saved 23, 32 and 42% of the total urine cultures at > 103, > 104 and > 105 CFU/mL respectively.
Comparison between a syphilis screen of two immunoassays for the diagnosis of syphilis
Saue E, Viia V, Mägi M
East-Tallinn Central Hospital Diagnostic Clinic, Central Laboratory, Tallinn, Estonia
Corresponding author: elo [dot] saue [at] itk [dot] ee
Background: Syphilis can be asymptomatic, serologic screening is recommended for persons at high risk, pregnant women, blood donors and routine survey. Treponema pallidum, the bacterium that causes syphilis, cannot be cultured. Serologic testing is treponemal method most often used to diagnose syphilis in patients with suspected disease. To evaluate the performance of the Immulite 2000 Syphilis Screen assay to the Cobas 6000 Treponema pallidum latex agglutination assay-TPLA. Parallel studies were carried out in order to switch to another method of analysis of the syphilis screening.
Materials and methods: Out of total 43 human serum specimens, belonging to three different categories (routine laboratory screening for syphilis, N = 16; syphilis patients, N = 15; potential false-positive results, N = 12), parallel studies were carried out in 16 unselected and 27 previously maintained positive serums. During the analysed period, parallel studies were performed by using the chemiluminescent immunoassay (Immulite 2000) and latex agglutination assay (Cobas 6000) method. All samples were tested with immunoblot Treponema IgG and IgM kits for confirmation.
Results: 6 results differed from the total number of 43 parallel studies carried out. 6 results were found to be false-positive since the confirmatory tests affirmed negative results. Syphilis screening can be switched over to the Cobas 6000 analyzer.
Conclusions: In comparison with the Syphilis Screen assay, the TPLA assay is more specific than the Immulite kit and has the advantage of reducing the number of confirmatory tests (mostly pregnant sera).
Procalcitonin as early diagnostic marker for infection in febril neutropenia cancer patients
Alonso T (1), Cuadrado-Cenzual M (2), De Gracia Y (2), Ortega De Heredia D (2), Perez-Segura P (1)
(1) Hospital Clinico San Carlos, Oncologia, Madrid, Spain
(2) Hospital Clinico San Carlos, Analisis Clinicos, Madrid, Spain
Corresponding author: mcuadrado [dot] hcsc [at] salud [dot] madrid [dot] org
Introduction: Febrile neutropenia (FN) in cancer patients is a complication related to antineoplasic therapy with serious impact on their morbidity and survival. The main cause is the infectious process. The aim of this study was to evaluate Procalcitonin (PCT) like early diagnostic marker in patient with FN associated with chemotherapy.
Materials and methods: A prospective double blind study was developed. We included 78 patients with FN and with treatment to them of the fever according to clinical practice protocol. Of all of them, 61 patients completed all the study with serial studies of PCT serum levels.
Results: Bateriemia was detected in 36 patients and PCT showed higher significantly levels (P < 0.001) in this patients in comparision with non bacteriemia patients. Relation between Procalcitonine values and treatment non–response was significant (p 0.999). The point of cut-off PCT level, with better sensitivity (74.6 %) and specificity (80%) was 0.21µg/L. The multivariant analysis showed that the value of PCT over 0.5 µg/L in patients with FN and bacteriemia it was an independent variable like marking diagnosis of bacteriemia in patients with febrile neutropenia (Odss Ratio 3.5 with an interval of 95% confidence 1.6-7.8) and P < 0.001.
Conclusion: PCT values in cancer patients with FN and infections; was higher and descend after restoring antibiotic treatment agreeing with clinical improvement. These data suggest it determination of PCT in cancer patients with FN; could be an useful early diagnostic marker for detection of bacteriemia.
The role of procalcitonin in sepsis patients with pneumonia
Cachapuz I (1), Amaro A (2), Granja C (2), Alves V (1)
(1) Matosinhos Local Health Care Unit, Clinical Pathology Service, Oporto, Portugal
(2) Matosinhos Local Health Care Unit, Intensive Care Medicine Service, Oporto, Portugal
Corresponding author: cachapuz [dot] isabel [at] gmail [dot] com
Background: The value of procalcitonin (PCT) in acutely ill sepsis patients has been studied and the use of this biomarker revealed its clinical relevance in ICU setting. The study purpose was to evaluate the role of PCT on septic patients management with community-acquired pneumonia (CAP) and invasive ventilator-associated pneumonia (IVAP).
Materials and methods: Prospective study. Adults admitted in ICU (April - December 2010) with CAP and/or IVAP diagnosis. PCT and C-reactive protein (CRP) were dosed daily. All patients underwent microbiological testing. Severity indexes were assessed. SPSS(v16) was used for statistical analysis.
Results: Total of 43 patients: 37 with sepsis related to CAP; 6 with sepsis related to IVAP. 74%male, average age:57 years old. 70% of the cases had a septic shock and 21% had severe sepsis. Blood cultures were positive in 11.6%; 25.6% had positive tracheal aspirate cultures; 26% were positive for antigenuria. Mean SOFA: 9 (SD 3.9);APACHE II: 20 (SD 8.5). Length of stay: 11.1 days. Mortality:14.7%. On admission day, PCT mean value: 23.7 ng/mL and CRP: 20.9 mg/dL. On day 1 serum PCT revealed a significant correlation with APACHE II index (P < 0.05) and was positive for blood cultures (P < 0.05).But, CRP value did not reveal these correlations. There was no relation either between PCT and SOFA index or with mortality.
Conclusions: This study revealed that even though PCT is related to APACHE II severity index, it is not related to ICU mortality. We found a significant correlation between PCT value and positive blood cultures, as mentioned in previous publications.
Extended spectrum Beta-lactamases and AmpC Beta lactamases producing uropathogenes among children
Deo P
Monash University, Department of Biochemistry and Molecular Biology, Melbourne, Australia
Corresponding author: pdeo2 [at] student [dot] monash [dot] edu
Objectives: UTI are common in childhood. Gram negative bacilli are the common isolates from the UTI. ESBL and AmpC beta-lactamase are the most significant enzymes involved in conferring resistance to Beta-lactam antibiotics in Gram negative bacteria. This study was aimed to find out the prevalence of multidrug resistant (MDR), ESBL and AmpC beta-lactamases producing isolates among children with UTI in Nepal.
Materials and methods: A prospective study was carried out from July, 2006 to March, 2008 at IOM,Kathmandu, Nepal). 820 urine specimens were obtained from clinically suspected UTI children (age < 12 years, female to male ratio 2.1:1). Most of samples were midstream urine, 25 supra pubic aspiration and 4 from catheter. Culture, organism identification and antibiotic susceptibility test were done by following the protocol of American Society for Microbiology (ASM).
Results: Among 820 urine samples, 23.51% (201/820) had significant bacterial growth with 184 (91.54%) non-repeat gram-negative isolates in which most were E. coli (58.15%) followed by Klebsiella species (15.2%).The prevalence of MDR, ESBL and AmpC were 115 (62.5%), 43 (23.36%) and 15 (8.15%) respectively. Maximum incidence of ESBL producer was E. coli (39.5%) followed by Klebsiella (16.2%) and Pseudomonas species (13.9%). Highly AmpC producing species were Klebsiella (40%) and Pseudomonas (26.6%). ESBL producers and non producers MDR isolates were highly resistant to amoxycillin-clavunic acid, aztrenome, cefepime and ceftazidime-clavunic acid. Imepenum and pipracilin were most effective drug among ESBL producers and non producers. Infection was more common in age group 6 ± 2.3 years with female to male ratio 2.04:1.
Conclusion: Result shows high percentage (62.5%) of MDR pathogens in childhood UTI.
Mass spectrometry in identification of antibiotic resistance
Pilatova K (1), Frostova T (1), Dolejska M (2), Budinska E (3), Sterba J (4), Zdrazilova Dubska L (1)
(1) Masaryk Memorial Cancer Institute, Department of Laboratory Medicine, Brno, Czech Republic
(2) University of Veterinary and Pharmaceutical Sciences, Department of Biology and Wildlife Diseases, Brno, Czech Republic
(3) Masaryk University, Institute of Biostatistics and Analyses, Brno, Czech Republic
(4) Clinic of Pediatric Oncology, Children's Medical Center, Brno, Czech Republic
Corresponding author: katerina [dot] pilatova [at] mou [dot] cz
Background: The assessment of antibiotic resistance is essential for appropriate treatment initiation of infection disease. Increasing frequency of multiresistant bacterial strains and development of new antibiotic resistance mechanisms represent major problems. Currently, mass spectrometry (MS) represents new approach for bacterial identification. The goal of this study is assessment of correlation between MS profile and bacterial antibiotic fenotype.
Materials and methods: We analyzed 56 isolates of Enterobacteriacea obtained from children treated for malignant disease. Bacteria were cultivated on agar plates; antibiotic susceptibility was tested using disc diffusion method, ESBL strains by double-disc synergy test. Sample preparation for SELDI-TOF MS: one bacterial colony was resuspended in 100 μL of distilled water and frozen at -70 °C. After thawing, suspension was applied on protein gold chip and after drying, covered twice by sinapinic acid matrix. Measurement was carried out on SELDI-TOF MS (Ciphergen) in the range of m/z = 3000 - 20000.
Results: We were able to clearly distinguish between individual bacterial species on the basis of cluster analysis of acquired protein spectra. We also found peaks which differ significantly in individual strains and correlates with antibiotic fenotype in Klebsiella pneumoniae.
Conclusions: Our pilot data show that protein profiling of bacteria using mass spectrometric methods allows discovery of new antibiotic resistance biomarkers and might represent new approach for identification of resistant bacteria strains as alternative method for current cultivation and molecular biology methods. This study was supported by European Regional Development Fund (RECAMO; CZ.1.05/2.1.00/03.0101) and by the Czech Science Foundation (grant P502/10/P083).
Prevalence of syphilis among blood donors in years 2009-2011 and our experiences with Abbott and Siemens reagents
Sopić T
University Clinical Centre of Maribor, Centre of Transfusion Medicine, Maribor, Slovenia
Corresponding author: tanja [dot] sopic [at] ukc-mb [dot] si
Background: Syphilis is sexually transmitted disease, caused by spirochaete organism Treponema pallidum. Although rare, it can be transmitted with blood transfusion and also is a good indicator of high risk sexual behaviours. Testing blood donors for syphilis is mandatory in most countries of the world.
Materials and methods: Specimens of blood donors were tested with Abbott reagents on Architect with CMIA (Chemiluminiscent Immunoassay) method and with Siemens reagents on BEP 2000 with EIA (Enzyme Immunoassay) method. Initially reactive samples were tested in duplicate. Repeatedly reactive samples were sent to confirmatory testing in reference laboratory.
Results: In years 2009-2011 we have tested 94,328 blood donations, 42,543 with Abbott reagents and 51,785 with Siemens reagents. There were 97 (0,23 %) initially reactive and then 93 (0.22%) repeatedly reactive with Abbbott reagents and 62 (0.12%) initially reactive and then 43 (0.08) repeatedly reactive with Siemens reagents. Confirmatory testing was positive for 17 (0.02 %) samples.
Conclusion: All confirmed positive results were reactive with Abbott and Siemens reagents, so both are suitable for screening for syphilis. We noticed more more false reactive results with Abbott reagents, then with Siemens reagents. The prevalence of syphilis among blood donors is staying the same in the last three years.
HBV pre-existing immunity of healthcare workers in occupational exposures in University Hospital Dubrava
Serdar T (1), Markovic D (2), Marijancevic D (1), Petrovecki M (3), Romic Z (1)
(1) University Hospital Dubrava, Clinical Department for Laboratory Diagnostic, Zagreb, Croatia
(2) University Hospital Dubrava, Department for Clinical Microbiology and Hospital Infections, Zagreb, Croatia
(3) Clinical Department for Laboratory Diagnostics, University Hospital Dubrava, Zagreb, Department of Medical Informatics, Rijeka University School of Medicine, Rijeka, Croatia
Corresponding author: tserdar [at] kbd [dot] hr
Background: Occupational exposure in healthcare workers represents every contact with a material that carries the risk of acquiring an infection during the working activities. Viral infections are the main blood transmitted infections and the most frequent are hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV).
Materials and methods: Since 2002, every reported occupational exposure of healthcare worker was registrated by the Committee for hospital infections of University Hospital Dubrava. Database among others contains qualifications of the exposed worker and his pre-existing immunity to HBV (aHBs-titer).
Results: In 10 year period of occupational exposures monitoring (from 2002 to 2011) 451 cases were reported. The majority of occupational exposures were reported by nurses or medical technicians (55.4%), followed by medical doctor resident (12.6%), medical doctor specialist (8.0%), cleaner (6.9%), laboratory technician (3.3%) and others (13.8%). In 59.4% of the exposed healthcare workers aHBs-titer status was assessed as satisfactory (aHBs > 100 IU/L), in 19.5% was not satisfactory (aHBs < 100 IU/L) and in 21.1% was not measured.
Conclusion: The 10-year follow-up of occupational exposures in healthcare workers in University Hospital Dubrava has shown an increase in the number of reported events. Implementation of preventive measures, such as universal precaution measures and HBV vaccination results a significant reduction in the incidence of HBV infection among healthcare workers. In many countries, the number of healthcare workers that underwent HBV vaccination usually does not exceed 65%, indicating that HBV vaccination is not applied in a sufficient number of healthcare workers.
Trends in antibiotics prescribed for bacterial pneumonia with higher CRP in preschool children
Bajraktarevic A (1), Kaljic Junuzovic A (1), Miokovic M (1), Penava S (1), Mulabegovic J (2), Delic M (3)
(1) Public Health Institution of Canton Sarajevo, Pediatrics department, Sarajevo, Bosnia and Herzegovina
(2) Pharmacy Faculty, Department for Biochemistry and Clinical Pharmacologz, Sarajevo, Bosnia and Herzegovina
(3) Krankenhaus Hietzing Stadt Wien Mit Neurologishem Zentrum Rosenhögel, Insitute Für Labormedizin Mit Serologie Und Infektiondiagnostik Mit Ambulanz , Vienna, Austria
Corresponding author: bajrakm [at] seznam [dot] cz
Objective: To estimate the incidence rates of preschool children outpatient bacterial pneumonia, examine time trends in antibiotics prescribed for bacterial pneumonia with higher CRP, and determine factors associated with broad-spectrum antibiotic prescribing for pneumonia in this population.
Materials and methods: The material consists of 902 small preschool Bosnians children aged 6 months to 6 years or half year more who took part in this retrospective study in pediatrics settings in six municipalities from nine municipalities of Canton Sarajevo. Within 72 or 96 hours after establition of diagnosis and beginning of antibiotics therapy blood samples CRP were taken for first and control analysis.
Results: The four most commonly prescribed antibiotic classes for bacterial pneumonia in preschool children were cephalosporins, macrolides, penicillins and aminoglycoside antibiotics. Cephalosporins were most commonly prescribed, ranging from 33.3% to 44.5% of all antibiotics prescribed for pneumonia in ages from 6 months to 6 years. Macrolides were the second most commonly prescribed antibiotic, ranging from 22.5% to 35.3% of all antibiotics prescribed for this diseases. There was no statistical difference in serum CRP values among the four groups with pneumonia after antibiotics therapy.
Conclusions: A strong association has been found between the level of circulating C-reactive protein (CRP) and the severity of pneumonia and success of antibiotics therapy in control laboratory data after three or four days in outpatients conditions .