H. De Beenhouwer

Detection of fastidious mycobacteria in human intestines by the polymerase chain reaction.

The aim of this study was to determine whether difficult-to-grow mycobacteria are present in human intestines. Intestinal tissue samples were subjected to both mycobacterial culture and a polymerase chain reaction (PCR) assay. After detection by PCR, species identity was determined by hybridizing the amplified 16S rRNA gene fragments with species-specific oligonucleotides. Intestinal biopsies from 63 patients with noninflammatory bowel diseases (n=22), Crohns disease (n=31), or ulcerative colitis (n=10) were analyzed. Culture and PCR revealed mycobacteria in four (6%) and 25 (40%) samples, respectively. Samples positive by PCR were negative with all probes specific to nine common cultivable species but were positive with theMycobacterium genavense-specific probe in 68% of cases. Mycobacterial isolates were identified asMycobacterium gordonae andMycobacterium chelonae. Findings were similar in Crohns disease samples compared to non-Crohns disease samples. This study shows that difficult-to-grow mycobacteria can be detected by PCR in large and similar proportions of inflamed intestinal tissue from patients with inflammatory bowel disease and intestinal tissue that appears normal from patients with noninflammatory bowel disease.

Atypical pneumonia due to Chlamydophila psittaci: 3 case reports and review of literature.

Abstract Chlamydophila psittaci is the causative agent of psittacosis or ornithosis. The disease is transmitted to men predominantly from birds. Most commonly noted symptoms are fever, headache and cough, but a number of other symptoms or complications may arise such as renal impairment, hepatitis or neurological symptoms. In this article 3 cases of psittacosis are presented, with a review of the literature with emphasis on laboratory diagnosis.

A hospital-level cost-effectiveness analysis model for toxigenic Clostridium difficile detection algorithms

BACKGROUND Despite thorough analyses of the analytical performance of Clostridium difficile tests and test algorithms, the financial impact at hospital level has not been well described. Such a model should take institution-specific variables into account, such as incidence, request behaviour and infection control policies. AIM To calculate the total hospital costs of different test algorithms, accounting for days on which infected patients with toxigenic strains were not isolated and therefore posed an infectious risk for new/secondary nosocomial infections. METHODS A mathematical algorithm was developed to gather the above parameters using data from seven Flemish hospital laboratories (Bilulu Microbiology Study Group) (number of tests, local prevalence and hospital hygiene measures). Measures of sensitivity and specificity for the evaluated tests were taken from the literature. List prices and costs of assays were provided by the manufacturer or the institutions. The calculated cost included reagent costs, personnel costs and the financial burden following due and undue isolations and antibiotic therapies. Five different test algorithms were compared. FINDINGS AND CONCLUSION A dynamic calculation model was constructed to evaluate the cost:benefit ratio of each algorithm for a set of institution- and time-dependent inputted variables (prevalence, cost fluctuations and test performances), making it possible to choose the most advantageous algorithm for its setting. A two-step test algorithm with concomitant glutamate dehydrogenase and toxin testing, followed by a rapid molecular assay was found to be the most cost-effective algorithm. This enabled resolution of almost all cases on the day of arrival, minimizing the number of unnecessary or missing isolations.

BRAIN ABSCESSES WITH PEPTOSTREPTOCOCCUS: NOT UNUSUAL AFTER OESOPHAGEAL DILATATION

Abstract A case of a brain abscess following oesophageal dilatation for caustic stenosis in a 67-year old woman is reported. Previously reported cases of brain abscess development after oesophageal dilatation are reviewed. Following oesophageal dilatation, bacteraemia and fever are common but the occurrence of metastatic brain abscesses is rare. The clinical presentation is non-specific, with high fever and neurological findings as most reported signs. The isolated organisms belong to the normal oropharyngeal bacterial flora. Prognosis is satisfactory after early diagnosis and correct management. As a result, clinicians dealing with oesophageal strictures should keep in mind that brain abscess formation is a potential complication of oesophageal dilatation.

Staphylococcus saprophyticus bacteremia after ESWL in an immunocompetent woman.

Abstract Staphylococcus saprophyticus is a well-known cause of uncomplicated urinary tract infections, especially in young and sexually active women. Presence in blood cultures is rare and often attributed to contamination. When bacteremia is significant, it occurs mostly in patients with hematologic malignancies and is predominantly catheter-related. However, we describe a case of significant bacteremia with S. saprophyticus associated with urinary tract infection after extracorporeal shock wave lithotripsy of an ureterolithiasis in an otherwise healthy patient.

The “Snotbarometer”: Epidemiological data on respiratory infections

no: 150 Presentation at ESCV 2016: Poster 184 The “Snotbarometer”: Epidemiological data on respiratory infections A. Vankeerberghen ∗, K. Dierickx, A. Boel, K. Van Vaerenbergh, H. De Beenhouwer Laboratory of Microbiology, OLVZ Aalst, Belgium Molecular detection of respiratory viruses was initiated in the Laboratory of Microbiology of OLVZ Aalst, Belgium, in 2003 with the detection of human metapneumovirus (hMPV) and respiratory syncytial virus (RSV). Since then, a constant elaboration of the portfolio was performed resulting in 8 multiplex in house real time PCR’s that detect 22 respiratory pathogens including viruses (RSV, hMPV, adenovirus, bocavirus, para-influenzavirus (PIV) 1, 2, 3 and 4, Influenza A and B, coronaviruses, enterovirus and rhinovirus) and atypical bacteria (M. pneumoniae, C. pneumoniae, B. pertussis, parapertussis and holmesii). Samples are mainly obtained from our hospital but also from other hospitals from the Flanders region. On each respiratory sample for which molecular diagnostics for at least one of these pathogens is requested, the complete PCR panel of 22 pathogens is performed. This increases the accuracy of a specific diagnosis, and it also results in “local” epidemiological data. These data are translated into a graphic representation, called the “snotbarometer”, which is made available for the hospital staff through the intranet, and on the website of the hospital. The “snotbarometer” consists of a weekly and a monthly report. In the weekly report, the amount of positive samples for each pathogen separately is depicted in a graph and updated weekly. This presentation gives the physician an idea of the actually circulating pathogens, of the amount of samples analysed in the lab, and the percentage of samples positive for each pathogen. In the monthly report a seasonal overview is given for the pathogens with epidemiological data available for multiple years, so one can start to extract the characteristic seasonal patterns. Examples are RSV, influenza A and B, PIV1, PIV2, PIV3 and PIV4. This year, Influenza B exceptionally preceded Influenza A which prolonged the influenza season. For other pathogens like adenovirus, bocavirus and M. pneumoniae the seasonality is less clear and one can observe a more fluctuating presence. Together, this information is very useful to predict the upcoming viruses. Conclusion: Regional epidemiological data are powerful since they can give useful information to the physician, especially when a weekly follow-up is available. http://dx.doi.org/10.1016/j.jcv.2016.08.224 Abstract no: 181 Presentation at ESCV 2016: Poster 185 Molecular characterization of human parainfluenza virus type 3 (HPIV-3) among hospitalized patients from central Israel I. Jornist 1,∗, E. Mendelson 1, D. Ram 2, R. Azar 2, M. Mandelboim 1, M. Hindiyeh 1 1 Chaim Sheba Medical Center & Tel-Aviv University, Israel 2 Chaim Sheba Medical Center, Israel Human parainfluenza virus 3 (HPIV-3) is an enveloped, non-segmented, negative sense RNA virus that belongs to the Paramyxoviridae family. HPIV-3 is a common cause of bronchiolitis and pneumoniae in children less than 1 year of age and one of the leading causes of acute lower respiratory tract infections in children under five years of age. In Israel, the epidemiology of HPIV-3 infections is not well characterized. In this study, epidemiology and molecular characterization of HPIV-3 was performed on patient samples collected between January 2012 and September 2015. Nasopharyngeal swabs (N = 15,946) were collected from hospitalized patients presenting with respiratory illness. Viral nucleic acid was extracted from patient sample using NucliSENS® easyMAG® (bioMérieux, France) and tested for the common human respiratory viruses (influenza viruses A and B, hMPV, adenovirus, RSV and HPIV-3) using validated real time PCR multiplex assays. Furthermore, molecular characterization of HPIV-3 complete HN gene (1722 bases) was performed after sequencing the complete HN gene. The Bayesian Markov chain Monte Carlo (MCMC) method was applied using a relaxed molecular clock, as implemented in the BEAST program (version 1.7.5). Trees were visualized and edited with the FigTree program (version 1.4.2) included in the BEAST software package. Of the patient samples tested, 547 (3.43%) samples were positive for HPIV-3. Stratifying HPIV-3, by month revealed the virus major activity was during the winter and spring seasons. Not only that, but the majority of patients infected were children less than 1 year of age and elderly greater than 60 years of age. An increased HPIV-3 activity was seen in patients hospitalized in the oncology/transplants wards of the hospital. Of interest were patient’s co-infections with HPIV-3 and other respiratory viruses. Of the 547 patient infected with HPIV-3, 99 (18.1%) patients were co-infected with other human respiratory viruses. Of which, adenovirus (6.6%) and RSV (6.4%) were the most common. Molecular characterization of the complete HPIV-3 HN gene from 50 different patients infected throughout the study period revealed that the majority of the HPIV-3 strains circulating in Israel belonged to the C1b and C3a clades. These HPIV-3 clades were mainly seen in the America’s and Saudi Arabia. In addition, one HPIV-3 isolate from the year 2012 did not match with any of the C1 clades, suggesting the possibility of being a new sub clade. HPIV-3 HN sequence analysis also revealed that the isolates characterized from Israel did not acquire the substitutions T193I and I567V in the HN gene suggesting that in patients with severe infection and where Zanamivir treatment is warranted, this antiviral can be used to help in managing the HPIV-3 infection. This is the first comprehensive study that characterized HPIV3 infections in Israel. The high co-infection rate of HPIV-3 and other common human patients mandates careful evaluation of the clinical presentation of infected patients and their prognosis. In addition, in depth evaluation of the clinical presentation of patients infected with the different HPIV-3 clades should be entertained. http://dx.doi.org/10.1016/j.jcv.2016.08.225

Respiratory viruses in the intensive care unit: More frequent than expected

no: 146 Presentation at ESCV 2016: Poster 151 Case report: Unexpected cause of respiratory failure 3 days after heart transplantation K. Dierickx ∗, A. Vankeerberghen, A. Boel, K. Van Vaerenbergh, H. De Beenhouwer Laboratory of Microbiology, OLVZ Aalst, Belgium Respiratory syncytial virus is an RNA virus belonging to the Paramyxoviridae and it is mostly found in young children. This virus can also cause morbidity and mortality in immunocompromised adults. Respiratory virus infection (RSV) is an important complication in solid organ transplant patients but the longitudinal monitoring of these infections has not been extensively studied. Little has been described in literature regarding RSV pneumonia in adult heart transplant patients. Here we report an interesting case of a 56 year old female with a history of non-ischemic cardiomyopathy starting in 2011. On January the 3rd of 2015 she successfully underwent a heart transplantation. Although there were no signs of respiratory disease at the time of hospitalization she showed respiratory insufficiency three days post-transplantation. In the microbiology lab each respiratory sample is cultured and when indicated screened for a panel of 22 targets detected in 8 inhouse RT-PCR multiplexes. This molecular panel covers the most important pathogens of viral respiratory infections and atypical bacterial pneumonia. The first respiratory sample of this patient was a bronchial aspirate taken three days post transplantation. The bacterial culture was negative but the sample tested positive for RSV-A with a high viral load (Ct value of 23). Follow up samples 15 days and 35 days post-surgery were still RSV positive although with decreasing viral load (Ct value of 25 and 28 respectively). Culture of respiratory samples showed the presence of Staphylococcus aureus only 10 days after surgery so RSV is most probably the primary cause of the respiratory disease. RSV was still detectable 1 month after transplantation which might be explained by the immunosuppressive treatment of the patient. The heart transplantation was performed during the RSV season. Some days before the surgery the lady had taken care of her young grandchildren so there indeed was a potential risk of community-acquired transmission. Conclusion: Without testing for viral pathogens no accurate diagnosis for the respiratory failure of this patient could have been made. Since screening of adult patients for viral pathogens is not common practice at the IC-unit, this case illustrates the added value of molecular screening when signs of respiratory failure arise in adult immunocompromised patients. http://dx.doi.org/10.1016/j.jcv.2016.08.191 Abstract no: 148 Presentation at ESCV 2016: Poster 152no: 148 Presentation at ESCV 2016: Poster 152 Respiratory viruses in the intensive care unit: More frequent than expected A. Vankeerberghen ∗, K. Dierickx, A. Boel, K. Van Vaerenbergh, H. De Beenhouwer Laboratory of Microbiology, OLVZ Aalst, Belgium In the Laboratory of Microbiology of the OLV Hospital in Aalst respiratory samples (n = 3500/year), received from multiple hospitals spread all over Flanders, are analysed on a daily basis by in house multiplex real time PCR for a panel of viral and bacterial pathogens. The panel includes adenovirus, bocavirus, human metapneumovirus (hMPV), respiratory syncytial virus (RSV), parainfluenzavirus (PIV) 1, 2, 3 and 4, Influenza virus A and B, enterovirus, rhinovirus, coronaviruses, Bordetella pertussis & parapertussis, Mycoplasma pneumoniae and Chlamydia pneumoniae. Before 2014, the majority of samples originated from children. The severe influenza epidemic in the winter season 2014–2015 made clinicians aware that viral infections in adults are not that innocent at all. Moreover, in the “Influenza season”, not only Influenza circulated but also other viruses were cause of severe disease. Correct identification of the pathogen is indispensable to administer or withhold therapy. As a consequence, the request for the real time PCR respiratory panel on samples from adult hospitalized patients increased. In order to calculate the frequency of these pathogens in adult critically ill patients, a retrospective study was performed for the period September 2014 to May 2016 including patients transferred to the coronary care unit (CCU) and the intensive care unit (ICU) because of respiratory failure. Respiratory panel results of samples, obtained in the window from 3 days before to 5 days after transfer to the CCU and IC units, were included. From the 126 samples, 44 samples were positive (34.92%) with 41 samples (93.18%) positive for a viral pathogen and 3 samples (6.82%) positive for a bacterial pathogen (1 M. pneumoniae, 1 C. pneumoniae and 1 B. parapertussis). None of the samples were positive for adenovirus or parainfluenzavirus. As expected, Influenza A virus (n = 14) and Influenza B virus (n = 8) were the most frequent and 1 patient had a co-infection of both viruses. No other co-infection was found. Surprisingly, rhinovirus (n = 8) was found to be the third most frequent viral cause of infection. hMPV and RSV are known to cause severe respiratory problems in infants and RSV infections have also been observed in the immunocompromised host. In our study, not only RSV (n = 5) but also hMPV (n = 7) was found frequently and caused very severe “Influenza-like” disease. We can conclude that viral infections are a common cause of respiratory problems in the intensive care unit and screening of these patients might be an important clue in diagnosis and correct treatment. http://dx.doi.org/10.1016/j.jcv.2016.08.192 Abstract no: 161 Presentation at ESCV 2016: Poster 153no: 161 Presentation at ESCV 2016: Poster 153 Multidrug-resistant cytomegalovirus infection in a pediatric stem cell transplantation patient T. Bauters 1,∗, L. Florin 2, V. Bordon 3, R. Snoeck 4, G. Andrei 4, S. Gillemot 4, P. Fiten 4, G. Opdenakker 4, G. Laureys 3, E. Padalko 2 1 Department of Pharmacy, Ghent University Hospital, Ghent, Belgium 2 Department of Clinical Chemistry, Microbiology and Immunology, Ghent University and Hospital, Ghent, Belgium 3 Department of Pediatric Hemato-Oncology and Stem Cell Transplantation, Ghent University Hospital, Ghent, Belgium 4 Rega Institute for Medical Research, Department of Microbiology and Immunology, KU Leuven, Leuven, Belgium Background: Cytomegalovirus (CMV), a member of the Herpesviridae family, is characterized by a lifelong latency in the host. Clinical presentations of CMV infection are minimal in immuno-

Establishing quality control ranges for temocillin following CLSI-M23-A3 guideline

Abstract Objectives: This study aimed to establish acceptable quality control ranges for temocillin disk diffusion tests and Etest® minimal inhibitory concentrations. Methods: According to Clinical and Laboratory Standards Institute (CLSI) guideline, a Tier 2 quality control study was performed and involves seven laboratories. Each of them tested 10 replicates of two quality control strains (Escherichia coli ATCC 25922 and E. coli ATCC 35218) on three different media lots and, for disk diffusion, two disk lots. Results: Proposed zone diameter quality control ranges were 12–25 mm for E. coli ATCC 25922 and 19–28 mm for E. coli ATCC 35218. Proposed Etest quality control ranges were 3–24 mg/l for E. coli ATCC 25922 and 2–6 mg/l E. coli ATCC 35218. Conclusion: Based on our results, we would advise the use of E. coli ATCC 35218 as QC strain for temocillin susceptibility testing and Etest because ranges obtained are narrower than with E. coli ATCC 25922 and do not overlap temocillin breakpoint.