Poster Competition BioMedica 2018
This year I had the great privilege of being the chairperson of the Poster competition at Biomedica. There were over thirty abstracts accepted for the post graduate competition, all of which reflected the high standard of research being carried out nationally in both our hospitals and research
centres. I was also delighted to be involved in the introduction of the under-graduate poster competition section to the conference this year. This is a fantastic addition to the competition as it facilitates the show-casing of the tremendous research that is being carried out by our under-graduate
medical science students from the three accredited colleges in the Republic of Ireland. We hope that this section will continue to grow over the next few years and welcome the submission of under-gradate research.
I, together with a panel of judges, had the difficult task of judging both the undergraduate and postgraduate posters. Each poster was reviewed and judged independently by each member of the panel. Overall everyone agreed that the standard was high and it was difficult to choose but a
consensus was reached for both categories and awards given at the closing ceremony to very deserving winners. It was fantastic to see the variety and high quality of research that is being carried by Medical Scientists, both at an undergraduate and post graduate level, being displayed over
the two days. I would encourage anyone who is involved in research to showcase it at every given opportunity. Remember you have put in the hours of hard work, make sure to let others see the results which are very often not only invaluable to your laboratory but also to others.
Dr Debbie Corcoran, Academy Council
First Prize Winner
NOVEL GENOTYPING OF CRYPTOSPORIDIUM PARVUM VIA HIGH RESOLUTION MELT ANALYSIS OF TANDEM REPEAT LOCI.
Topic: Medical Microbiology
Authors: O’ Leary, JK(1); Blake L(2), Corcoran D(2), Sleator, RD(1, 3); Lucey, B(1)
1 Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Cork, Ireland
2 Department of Medical Microbiology, Cork University Hospital, Wilton, Cork, Ireland
3 APC Microbiome Institute, University College Cork, Cork, Ireland
Microscopy still provides the gold standard means by which to diagnose clinical Cryptosporidium infection. However, differentiation between species and sub-species for epidemiological purposes requires the use of molecular techniques, including real time PCR. There is currently no
consensus on the most suitable method for sub-speciation of Cryptosporidium spp1.
High resolution melt (HRM) analysis has been used successfully with pathogens other than Cryptosporidium for subtyping purposes2, while within Ireland, approximately 80% of clinical cases of Cryptosporidium infection are attributable to Cryptosporidium parvum3. The purpose of the current study was to develop a method capable of differentiating between C. parvum sub-species.
This was approached by first identifying a panel of established variable number tandem repeat (VNTR) loci known to vary between C. parvum sub-species. These VNTR loci were targeted via nested real time PCR and HRM analysis. HRM
analysis is based upon the detection of small genetic variations via minute differences in melting temperature determined when amplified double-stranded DNA is subjected to denaturation and reannealing. The generated HRM data was subsequently analysed via melting curves and melting peaks, allowing for differentiation between sub-species. Multi-locus sequence typing (MLST) analysis was employed in conjunction with HRM
in order to definitively identify the sub-type of each clinical sample and verify HRM results.
This application was employed to discriminate between allelic differences within the selected loci, based on the variable number and sequence of the tandem repeats within these regions. Our newly-developed method was tested using a set of 12 sequenced reference samples from the Cryptosporidium Reference Laboratory (CRU), Cardiff, Wales, and a bank of clinical Cryptosporidium-containing samples (2015-2018) from the department of Clinical Microbiology, Cork University Hospital. Our study reports on the analysis of these samples, and provides a novel method for the subtyping of C. parvum.
1. Chalmers RM, Robinson G, Hotchkiss E, et al. Suitability of loci for multiple-locus variable-number of tandem-repeats analysis of Cryptosporidium parvum for inter-laboratory surveillance and outbreak investigations. Parasitology. 2017;144(1):37-47.doi:10.1017/S0031182015001766
2. Keeratipibul S, Silamat P, Phraephaisarn C, et al. Genotyping of Salmonella enterica Serovar Typhimurium Isolates by Multilocus Variable Number of Tandem Repeat High-Resolution Melting Analysis (MLV-HRMA). Foodborne Pathog Dis. 2015;12(1):8-20. doi:10.1089/fpd.2014.1761
3. Zintl A, Proctor A., Read C, et al. The prevalence of Cryptosporidium species and subtypes in human faecal samples in Ireland. Epidemiol Infect. 2009;137(2):270-277. doi:10.1017/S0950268808000769
TARGETING THE DNA DAMAGE RESPONSE IN CANCER: FUNCTIONAL CHARACTERISATION OF NOVEL RAD9-INTERACTING PROTEINS
Kiely, AK(1), O’Halloran, FO’H(1), Grenon, MG (2,3), Lowndes, NL (2,3) and Finn, KF(1).
1. Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Co. Cork
2. Department of Biochemistry, School of Natural Sciences, National University of Ireland, Galway
3. Centre for Chromosome Biology, Biomedical Sciences Building, National University of Ireland, Galway
“By 2020 it is estimated that 1 in 2 adults will get a cancer diagnosis in their lifetime”. Cancer is a disease caused largely by changes to genes that control cell growth and proliferation. These genetic changes can be inherited and/or acquired by direct damage to DNA via exposure to DNA damaging agents, such as ultraviolet (UV) radiation and carcinogenic agents, as well as by-products of normal cellular metabolism. Cells have
evolved a complex system, termed the DNA damage response (DDR), which detects DNA damage and promotes DNA repair(1).
Rad9 was the first DNA damage checkpoint protein identified in budding yeast. Loss of Rad9 function results in significant sensitivity to DNA damaging agents due to defects in the DDR (2). In undamaged cells, Rad9 exists as a large complex (>850kDa) and is remodelled following DNA damage (3). A preliminary proteomic study has identified 55 putative Rad9-interacting proteins.
This project aims to phenotypically characterise these putative interactors to determine whether these proteins play an important role in Rad9-dependent DNA damage checkpoint activation and/or DNA repair. To test this hypothesis deletion strains will firstly be analysed via droptest to determine DNA damage sensitivity to a variety of DNA damaging agents.
To date several deletion strains screened have exhibited sensitivity to (UV) radiation (80Jm-2) and/or the double-stand break-inducing antibiotic, Zeocin (4Ìg/ml). Sensitivity to Hydroxyurea (HU)-mediated DNA damage is currently ongoing. Sensitive strains identified in the screen will be further characterised to determine whether the protein of interest plays a role in DNA damage checkpoint activation (G1, intra-S, G2/M) and/or DNA repair.
As cellular events governing the DDR are highly conserved from yeast to mammals, novel findings in budding yeast may be extrapolated to humans. Thus, data from this project could significantly impact cancer research and influence the development of novel cancer therapeutics.
1. Jackson SP. UKPMC Funders Group The DNA-damage response_: new molecular insights and new approaches to cancer therapy. Tennis.
2. Weinert TA, Hartwell LH. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae. Science .1988 Jul 15 [cited 2018 Mar 1];241(4863):317–22.
3. Gilbert CS, Green CM, Lowndes NF. Budding yeast Rad9 is an ATPdependent Rad53 activating machine. Mol Cell. 2001 Jul [cited 2018 Mar 1];8(1):129–36.
SUCCESSFUL IMPLEMENTATION OF DEMAND MANAGEMENT OF VITAMIN B12 AND FOLATE REQUESTS IN THE PATHOLOGY LABORATORY IN NAAS GENERAL HOSPITAL
Claire Burke, Chief Medical Scientist, Haematology Department, Naas General Hospital.
Authors: Claire Burke, Johnny McHugh, Ronan Desmond.
In the current economic climate it is vital that Laboratories play an increasing role in ensuring that testing is evidence based.
With this in mind, we at Naas General Hospital looked at B12 and Folate requesting patterns in our catchment area. In 2016, our laboratory received 55,077 requests for B12, Folate and Ferritin. This was an increase of 44% since 2011. Naas General Hospital provides B12, Folate and Ferritin testing for a population of 180,011, which is an increase of 6.9% since 2011(HSE health atlas). The increase in numbers doesn’t correlate with the population increase.
In May 2016 the National Laboratory Handbook Volume 1 was published and together with a GP survey that was sent to our GP’s, we decided to implement demand management for B12 and Folate test requests. The demand management process was implemented in June 2017.This process involved rejection of tests based on inappropriate clinical details and assessment of the patients full blood count. The
average monthly referral decreased by 48% in 2017 in the post implementation period. To determine whether the reductions in the number of total B12 samples processed and low B12 results were statistically significant we performed and independent- samples t-test.
By the end of 2017 there was a cost saving of at least €87,000 euro.
The issuing and enforcement of guidelines produced a dramatic reduction in testing without affecting the ability of Clinicians to detect clinically relevant B12 deficiency.
1. Freyer AA, Hanna FW. Managing demand for pathology tests: financial imperative or duty of care? Ann Clin Biochem 2009;200946:435-7
2. National Laboratory Handbook Volume 1
Undergraduate Poster Winner
INVESTIGATION OF THE OVERESTIMATION OF PARAPROTEINS IN SERUM BY IMMUNOTURBIDIMETRIC METHOD: TIME FOR A
CHANGE OF PROTOCOL IN SLIGO UNIVERSITY
Location: Sligo University Hospital & Galway-Mayo Institute of Technology
Authors: Roe, M., Mahon, C., Montgomery, N., McGrath, M.
A key aspect of the treatment and monitoring of patients with monoclonal gammopathies relies on obtaining reproducible and accurate quantitation of paraproteins. Discrepancies between quantitation of paraproteins by Serum Protein Electrophoresis (SPE) and Immunoturbidimetry has been well documented [1, 2]. In Sligo University Hospital (SUH), all immunoglobulin results are assayed and reported on a daily basis, independently of SPE results which may not be reported for some days later. The aim of this research was to: • Examine the extent of discrepancies that sometimes occur in the quantitation of a paraprotein band by SPE and the result of the immunoglobulin isotype by
• Evaluate the current practices in the reporting of SPE and immunoglobulin results particularly in patients with a paraprotein band
• Suggest recommendations (if required) for a change in the reporting protocol
A retrospective search was carried out of 240 serum samples that had paraproteins to examine the relationship between SPE and immunoturbidimetric analysis. Due to noted discrepancies, a selection of samples (n=75) were analysed for Immunoglobulins by Immunoturbidimetry and diluted if outside the reference range and reanalysed. From these samples, SPE (n=24) were performed. Identification of paraprotein bands was carried out and paraprotein concentrations quantified. Comparisons were made with the relevant immunoglobulin
Significant discrepancies between Immunoturbidimetry and SPE were evident, especially at higher paraprotein concentrations. Dilution of the samples did not offer an improvement in the overestimation of paraproteins by immunoturbidimetric analysis.
A consistent approach is required by laboratories and clinicians so that the paraprotein concentration is monitored on an individual basis using the same method. Based on the findings of this research project many recommendations were suggested. At a minimum, if immunoglobulin results are to be reported independently of SPE results, a disclaimer highlighting the possibilities of inaccuracies should be appended to the
result. Ideally, all immunoglobulin results should be withheld and reported only when SPE results are available. In cases where a discrepancy is evident, overestimated values by immunoturbidimetry should not be reported and only the non-paraprotein serum immunoglobulins reported to
demonstrate immunoparesis . In these cases the paraprotein concentration quantified by SPE should be given reported solely.
Update: These recommendations were implemented in SUH in January 2018. All immunoglobulin results (IgG, IgA, and IgM) outside the reference range are placed in a hold file and are validated in conjunction with SPE results.
 Jelinek, A.G. and Bachmann, L.M. (2014). Unexpected test results in a patient with multiple myeloma. Clinical chemistry, 60 (11), 1375-1378.
 Keren, D.F. and Schroeder, L. (2016). Challenges of measuring monoclonal proteins in serum. Clin Chem Lab Med, 54 (6), 947-61.
 Tate, J., Caldwell, G., Daly, J., Gillis, D., Jenkins, M., Jovanovich, S., Martin, H., Steele, R., Wienholt, L., Mollee, P., (2012). Recommendations for standardized reporting of protein electrophoresis in Australia and New Zealand. Annals of Clinical Biochemistry, 49 (3), 242-256.
Academy Conference Bursary Award
Carol Troy, Surveillance Scientist, Microbiology Laboratory, University Hospital Waterford
Iwas honoured to receive the Academy Conference Bursary Award this year. This award provided me with an
opportunity to present the findings of my Master’s thesis as a poster at the recent European Congress of Clinical
Microbiology and Infectious Diseases (ECCMID) conference in Madrid. The title of my thesis was “A Retrospective Review of Community Associated and Healthcare Associated ESBL Producing E. coli and K.
pneumoniae in the South East of Ireland 2015-2016” and was undertaken as part of a Masters in Healthcare Infection Management in Trinity College, Dublin. This review demonstrated a significant increase in ESBL producers over a 24 month period. High levels of resistance were observed among both healthcare associated and community associated isolates. The majority of isolates were found to be community associated indicating a reservoir of ESBL in the community.
ECCMID has been an annual event since 1999. It is the annual conference of the world’s largest society European Society for Clinical Microbiology and Infectious Diseases, (ESCMID) covering microbiology and infectious diseases. This year’s conference was held in Madrid from the 21st to the 24th of April. The programme of events over the four days included educational workshops, oral sessions, keynote lectures, meet-theexpert sessions and poster presentations. In addition to the extensive educational opportunities, a large arena was also dedicated to displays by leading diagnostic and pharmaceutical companies on new and emerging trends in laboratory diagnostics and drug therapies.
Being able to present my poster at the conference allowed discussion of my findings with fellow attendees and also to explore the numerous other areas of research and study currently being undertaken on ESBL producing organisms.
With a vast array of topics on offer over the course of the four days in the various lecture halls and presentation rooms, trying to get to see all my chosen lectures was a challenge!
As my job is primarily focused on surveillance my choice of lectures were primarily in this area. Some of the more interesting lectures I was able to see included Surveillance and Prevention of Surgical Site Infection and The Burden of Healthcare Associated Infection(HAI). The implementation of a SSI programme results in positive patient outcomes and can be achieved even in the resource limited environment as was demonstrated
by a study at a Tanzanian hospital. The speakers on HAI covered a variety of topics including ESBL HAI in ICU adding to the costs of isolation, but the overwhelming message was that HAI is a worldwide problem that is being fought in every healthcare environment.
Among the diagnostic companies’ stands that I visited, the ever growing emphasis on rapid identification and antimicrobial susceptibility testing was evident. In particular the area of bacterial identification where molecular testing is providing diagnostic answers at an ever increasing pace. This can only impact positively on both the patient and the healthcare environment allowing earlier treatment of patients, and identification of patients who are infected or colonised with MDROs.
I am extremely grateful to the Academy for awarding me this bursary that allowed me to bring the findings of my study to a wider audience and also to engage in the educational experience of ECCMID.
A Retrospective Review of Community Associated and Healthcare Associated ESßL Producing E.coli and K.pneumoniae in South East Ireland 2015-2016
Background: Extended-spectrum beta-lactamases (ESBL) are a major public health problem. A retrospective study was undertaken of ESBL-producing E. coli and K. pneumoniae isolated from clinical specimens in the regional diagnostic microbiology laboratory in the South East of
Ireland during the period January 2015 to December 2016. The aim of this study was to review the patient demographics, geographical data, primary acquisition site and antimicrobial susceptibility data.
Materials/methods: Data on all ESBL-producing E. coli and K. pneumoniae for 2015 and 2016 was extracted from the laboratory and patient information management systems. This included antimicrobial susceptibilities and patient demographics. Isolates were designated healthcareassociated, community-associated, or long-term care facility associated. Under these designations, data was analysed under the following headings: age and gender distribution, length of stay for patients admitted to an acute healthcare facility, specimen types, and antimicrobial susceptibilities. In total 75 isolates from the study were examined for the presence of blaCTX-M by real time PCR.
Results: A total of 896 ESBL-producing isolates met the study criteria; 822 E. coli and 74 K. pneumoniae. SBLproducing E. coli isolates increased from 255 in 2015 to 567 in 2016 and ESBL-producing K. pneumoniae isolates increased from 27 in 2015 to 47 in 2016. This represents an overall increase of 118% in ESBL-producing isolates. Community-associated isolates increased from 57% in 2015 to 67% in 2016, healthcare-associated isolates also increased from 15% in 2015 to 20% in 2016. Long-term care facility associated isolates decreased from 28% in 2015 to 13% in 2016. Female patients represented 73% of isolates and 71% of patients >65 years of age. Urine accounted for 730 of the 896 specimens. The proportion of isolates that were multidrug-resistant rose from 74% in 2015 to 82% in 2016. In total 70/75 (90%) harboured a blaCTX-M gene (63 = blaCTX-M group 1, 7 = blaCTX-M group 9)
Conclusions: Community-associated ESBL producing E. coli and K. pneumoniae represent an increasing threat to public health. Failure to control the spread of these isolates through prudent antimicrobial use and screening may potentially result in an increase in the use of carbapenems and the spread of carbapenem resistance.