Public Health Wales Research and Development 2014/15 The following table contains all research and development projects which concluded in the 2014/15 financial year. # 1 Project Title Characterisation of HPV viral integration in DNA obtained from residual cervical smears collected within Baseline HPV 2009 Lay Summary Human Papillomavirus (HPV) is a very common viral infection and most women will be infected at some point in their lives. This is a study to see if carrying out new tests for HPV status on cervical screening samples could identify women at a higher risk of cervical disease development. Occasionally, HPV can cause abnormal cell growth by interfering with the normal cell controls. Public Health Wales Investigator Helen Beer (Senior Information Manager and Research Specialist, Screening Division) Helen.Beer@wales.nhs.uk The tests will start to look for possible changes in the virus status that could be the cause of this loss of control. This study will be performed on stored residual smear material (80°C) that has been previously collected as part of a pseudoanonymous study and will not affect patient treatment and will not require additional samples. 2 KESS Project: Developing an integrated whole genome amplification and next generation High risk (HR) HPV positive samples will be tested further with for viral integration status. The results of these tests will be compared with available smear and biopsy results collected pseudo-anonymously over 2 rounds of screening (up to 6 years) as part of the original Baseline HPV 2009 study. Cryptosporidium is a group of protozoan parasites which cause an intestinal disease called Cryptosporidiosis. Infected people develop symptoms such as diarrhoea and intestinal pain but usually get better within a few weeks. However, in patients with weak immune systems the infection can be fatal. There 1 Professor Rachel Chalmers (Consultant Clinical Scientist/Honorary Professor Head, Cryptosporidium Reference Unit Public Health Wales Microbiology) Useful Links and Publications sequencing method to characterise Cryptosporidium clinical isolates is no licensed treatment for cryptosporidiosis. To rachel.chalmers@wales.nhs.uk develop better treatments, understand how the disease spreads and how it can be prevented researchers are studying the DNA of the parasite by so-called “next generation sequencing” (NGS) techniques. For NGS to be successful, enough parasite DNA needs to be extracted from patient stool samples. However, often only small amounts of parasite DNA can be extracted, and NGS is not possible. To be able to analyse samples containing small amounts of parasite DNA, we want to use a method called “whole genome amplification” (WGA). This is a way to make many copies of a DNA sample and will hopefully make enough copies of the parasite DNA in samples with amounts to allow us to analyse the sequence by NGS. The first aim of the project then is to test whether WGA and NGS, in combination with other preparation techniques, works by analysing human clinical samples. The second aim of the project is to use the method to analyse the DNA sequences of further clinical isolates to investigate the Cryptosporidium species causing outbreaks and sporadic disease in Wales and the UK and to help develop better methods for identifying outbreaks of cryptosporidiosis. The stool samples used in the study will be selected from those received at the Cryptosporidium Reference Unit (CRU), Public Health Wales Microbiology, Singleton Hospital, Swansea for DNA analysis by routine DNA typing methods and anonymised before testing. The study will be undertaken at the CRU and Aberystwyth University. 2 3 4 A survey to investigate the reasons why patients in the ‘standard’ and ‘nonurgent’ triage categories attend the Emergency Department (ED) and to explore what other options they have considered. Development and assessment of a Nucleic Acid Amplification Test for the detection of Toxoplasma gondii in meat entering the human food chain The ultimate aim of the research is to investigate the reasons why patients in the ‘standard’ and ‘non-urgent’ triage categories attend ED and to explore what other options they have considered before attending the ED. An anonymous self-completion questionnaire will be designed, pre-piloted and then produced in both English and Welsh. This will be in a format to allow electronic analysis, working closely with Bangor University. The survey will also collect basic demographic information on patients (age, sex and postcode). The setting will be the 3 EDs in North Wales and the survey will be conducted in February and March of 2014. The survey instrument will be administered by Betsi Cadwaladr University Health Board (BCU HB) Emergency Department staff in the 3 EDs to patients and/or their carers (aged 16 years+) post-triage; this will be limited to patients who have been flagged up as being in the ‘standard’ and ‘nonurgent’ triage categories (Manchester triage categories green and blue). Those patients who are incapable through alcohol or drugs to complete a questionnaire will be excluded from the survey, as well as those who are in emotional distress, or those who have been referred to the ED formally by letter from their GP. Toxoplasma gondii is a parasite which affects humans and can cause ocular toxoplasmosis, congenital toxoplasmosis and miscarriage/stillbirth. The parasite is transmitted via ingestion of the oocysts through the consumption of undercooked meat, ingestion contaminated water or from mother to foetus. The purpose of this study is to develop and assess a Nucleic Acid Amplification Test for the detection of oocysts in 3 Dr Robert Atenstaedt (Consultant in Public Health Medicine, Public Health Wales) Robert.Atenstaedt@wales.nhs.uk Professor Edward Guy (Consultant Clinical Scientist - Head of Toxoplasma Reference Unit, Public Health Wales) Edward.guy@wales.nhs.uk Atenstaedt, R, Gregory, J, PriceJones, C, Newman, J, Roberts, L, and Turner, J. (2014). Why do patients with nonurgent conditions present to the Emergency Department despite the availability of alternative services? Epub ahead of print 17 November. In: Eur J Emerg Med :pp. 5 Evaluation of the Bactec fx system in combination with various novel molecular methods for the detection and identification of Candida species causing Candidaemia mixed meat and ham. The results will provide a guideline for future detection of toxoplasma in meat. This project will focus on determining the threshold of BACTEC fx to detect candidaemia, the most common infestation of invasive candidiasis. Currently the sensitivity is 50% and is due to the low organism numbers in circulation and consequently, collecting blood samples, resulting in a necessity for a higher frequency of testing to increase the chances of detection. Growth also regularly takes a number of days in blood cultures, with extra time then required for conventional phenotypic identification. Identification is required for effective treatment as certain species are Azole resistant. A range of Candida species will be inoculated at different CFU/ml levels into different BACTEC bottle types and the time for a bottle to become positive will be monitored and extended if the culture remains negative after 5 days with end plating of all broth being done to determine the threshold of the BACTEC fx. Detection and identification methods ranging from chromogenic agar, matrix-assisted laser desorption ionization-time of flight (MALDI TOF) mass spectrometry, real-time polymerase chain reaction (PCR) and Raman spectroscopy will be used directly on blood cultures and cultured isolates to determine the ID of the various Candida species. The use of rapid and sensitive techniques, if shown to be reliable, could hopefully speed up the diagnostic process, reduce mortality rates and allow appropriate antifungal treatment to be given to those individuals with invasive 4 Paula Brookes (Operational Manager Bacteriology, Public Health Wales) PaulaBrookes2@wales.nhs.uk 6 7 What is the unrecognised UK prevalence of Coxiella Burnetti in patients prior to cardiac Valve replacement surgery: A pilot study disease and reduce unnecessary therapy and toxicity in other patients, resulting in cost saving for the NHS. Q fever is an uncommon disease that in some cases can cause chronic damage to the valves of the heart. This often happens with very few symptoms and is often difficult to differentiate from age related change to the heart valves. There has not been a study before in the UK of how many people who need new heart valves may have Q fever without knowing it. Dr Brendan Healy (Consultant in Microbiology and Infectious Diseases, Public Health Wales) Brendan.Healy@wales.nhs.uk We aim to study a group of people undergoing routine heart valve replacement and to test them for serological (blood) evidence of Q fever and to test the valve that is removed during their operation for Q fever. All these tests will happen while the patient is asleep during their operation. We aim to use the results to estimate the number of people undergoing routine valve replacement that have evidence of Q fever infection and then see how that may affect the lifespan of their new valve. Understanding In Wales, bowel screening is currently offered to men healthcare and women aged between 60 and 71 years old. utilisation of Screening programmes require good uptake to be responders and non- effective. In order to develop a strategy for increasing responders uptake we first need to find out how non-responders to the bowel differ from responders. For bowel screening, which is a screening test undertaken at home, there may be opportunities to programme in Wales promote uptake through contact with categories of before and after health professionals not usually associated with the invitation to bowel screening programme. Through understanding screening in order to healthcare utilisation we hope to find potentially useful improve early intervention points and conversely, and not waste time 5 Dr Rosemary Fox (Director of Screening Division, Public Health Wales) Rosemary.fox@wales.nhs.uk http://medicine.cf.ac.uk/primarycare-publichealth/research/healthyageing/completedprojects/understandinghealthcare-utilisationresponders-and-non-responde/ detection either through screening or opportunistic testing 8 A prospective study of faecal parasite infections among children in Cambodia and resources pursuing strategies unlikely to be successful. This study aims to compare the engagement of responders and non-responders to the bowel screening programme with health care services in the year before and the year following invitation. We will explore the engagement with primary and secondary care before and after the screening invitation in order to compare responders and non-responders and to identify possible strategies for encouraging participation. We plan to link information from several sources to compare morbidity, measured by GP consultations, hospital outpatient, elective and emergency admissions, both before and after invitation to the screening programmes. Cambodia has some of the poorest health indicators in Southeast Asia, with high rates of malnutrition. Just under a third of the population have access to improved sanitation facilities, and rainwater is the main source of water for most homes. It is therefore not surprising that infection with gut parasites is a common problem in children, causing detrimental effects on nutrition, growth and cognitive development and contributing substantially to childhood anaemia. The primary purpose of this study is to define the incidence and range of faecal parasites causing infections in Cambodian children presenting to hospitals; the R&D approval sought here is specifically for the work that will be undertaken at the national Cryptosporidium Reference Unit in Public Health Wales Microbiology Swansea: the analysis of DNA extracted from stools for 6 Professor Rachel Chalmers (Consultant Clinical Scientist/Honorary Professor Head, Cryptosporidium Reference Unit Public Health Wales Microbiology) Rachel.chalmers@wales.nhs.uk the detection and characterisation of the Cryptosporidium parasite. The DNA will have already been extracted from stools at Liverpool University. The National Public Health service for Wales will shortly be rolling out a new form of test (called a dried blood spot test) for hepatitis B, hepatitis C and HIV infection amongst individuals at high risk of infection. The dried blood spot test involves few drops of blood being taken by a safe and simple finger prick device. The dried blood spot test samples will be taken by substance misuse services and specialist nurses and tested by the Welsh specialist virology unit. The study aims to see how good the new test is when compared to the previously used test (a blood sample taken with a needle and syringe) for detecting hepatitis B, hepatitis C and HIV infection. This potentially allows the test in future to be offered as a first line diagnostic tool rather than at present as a ‘pre test’ requiring further confirmation. Individuals coming forward for routine diagnostic blood borne virus testing by a blood sample taken with a needle and syringe will be asked to provide a dried blood spot sample as well. The results of the two tests will be compared allowing an assessment of the quality of the dried blood spot test to accurately detect an infection. 9 Sensitivity and specificity of dried blood spot testing for the diagnosis of blood born viral infection 10 Identification of antibacterial components of oils and herbs previously found to have antibacterial activity In a previous study it was shown that oils and herbs showed antibiotic like properties against certain gut bacteria. The purpose of this study is to look at the oils and herbs constituent components to see if there are specific constituents of these compounds that confer the antibiotic activity. 11 A prospective randomised, Blood stream infections (BSI) are a major problem and Dr Robin Howe (Consultant about 8% of hospital inpatients have a BSI. Some will be Microbiologist, Public Health 7 Dr Rachel Jones (Consultant Virologist, Wales Specialist Virology Centre, Public Health Wales) rachel.jones11@wales.nhs.uk Eugene Rees (Technical Head of Bacteriology, Public Health Wales Microbiology Swansea) Eugene.rees@wales.nhs.uk multicentre trial to assess the impact of laboratory based RAPId Diagnosis on Outcome in patients with Blood Stream Infections (RAPIDO) admitted with a BSI and others will develop an infection Wales) during their hospital stay. Recent improvements in Robin.howe@wales.nhs.uk infection control have reduced the number of patients that acquire an infection during their hospital stay, but BSI still occur and there are currently around 90100,000 cases a year. In certain infections the death rate can be as high as 50% and every patient that contracts an infection has a longer hospital stay. BSI is diagnosed by taking a blood sample from patients with suspected infection. Antibiotic treatment is commenced within hours, but processing of blood cultures takes several days to reach a definitive diagnosis and identify the correct antibiotic. The initial antibiotic selected will be a broad spectrum antibiotic, based on the clinician's judgment of the clinical picture. However, until definitive bacterial analysis is performed it is unknown if the correct antibiotic has been selected and there can be a time lag of up to 35 days before patients receive appropriate antibiotic therapy for their BSI. Administration of the correct antibiotics has been shown to reduce death by up to 50% in some patient groups. This study will assess the impact of new technology designed to speed up laboratory diagnosis. The new technique should produce information within ½ hr of commencing laboratory processing of positive specimens. A five centre, randomised controlled trial design will be used and all patients with BSI will be randomised into two groups, one which will be subjected to the current diagnostic approach and the other group will have the 8 12 13 Human Papillomavirus integration analysis in liquid based cytology samples as a biomarker for cervical disease A pilot study on the current diagnostic approach plus the new rapid diagnostic technology as well. Outcome will be assessed by collecting the same set of clinical information on each patient in the study. This is a study to see if carrying out new tests on cervical screening samples could safely reduce the number of women referred to hospital following an abnormal smear result. Taking part in the study will involve having one additional smear taken but will not affect patient treatment. Identifiable personal details will be removed from the samples before they are tested. The result of the tests will only be used by the study team to evaluate the new tests for viral integration and host biology markers. The result will not be included in the women’s hospital or screening records. The women will be invited to take part if they have been referred to the colposcopy clinic following an abnormal smear. If they consent to being included in the study, a smear will be taken and tested for human Papillomavirus (HPV) before colposcopy. Helen Beer (Senior Information Manager and Research Specialist, Screening Division) Helen.Beer@wales.nhs.uk HPV is a very common virus that is responsible for cervical cancer. HPV types 16 and 18 are considered to be high risk and, together with type 45, are the three commonest HPV types in cervical cancer, causing about 80% of cervical cancers. HPV 16, HPV 18 and HPV 45positive samples will be tested further to examine how far the virus has integrated into the cervix. The results of these tests will be compared with available smear and biopsy results from consenting women at subsequent visits (up to 4 years). These results will be provided by Screening Services Wales Toxoplasma is a common single-celled parasite that Professor Edward Guy (Consultant 9 risk factors for Toxoplasma Gondii infection in England and Wales infects about 5 in 1000 of the UK population each year. Once an individual is infected, the parasite forms cysts in heart and CNS tissues and can remain there for life. In otherwise healthy individuals Toxoplasma infection often passes unnoticed or causes only mild to moderate flu- or glandular fever-like symptoms that require no treatment. If the infection is acquired during pregnancy, however, this can be severe or lifethreatening for the unborn child. Clinical Scientist - Head of Toxoplasma Reference Unit, Public Health Wales) Edward.guy@wales.nhs.uk Toxoplasma is acquired either by eating raw or undercooked meat from animals that are infected, or by inadvertently ingesting oocysts (microscopic parasite eggs) from the environment. Thus, infection may be food borne or may be associated with a wide range of occupational or social activities involving exposure to the environment. In the UK it is not known which of these routes is the most important and this information is critical in ensuring health protection advice is targeted towards the highest risk activities. The findings will also inform national strategy and policy on food safety. 14 Role of chronic urinary infection in the pathophysiology of overactive bladder syndrome This study involves finding out, through a 30 minute telephone interview and questionnaire, what exposure to risk factors has been encountered by individuals recently infected with Toxoplasma. Overactive bladder (OAB) is characterised by urgency (a sudden compelling desire to pass urine which is difficult to defer), with or without urge incontinence (complaint of involuntary leakage accompanied by or immediately preceded by urgency), usually with frequency and nocturia (complaint that the individual has to wake at 10 Eugene Rees (Technical Head of Bacteriology, Public Health Wales Microbiology Swansea) Eugene.rees@wales.nhs.uk night one or more times to void).It effects 13% of the population and the incidence rises with age. It has significant adverse effect on quality of life; sleep quality and emotional wellbeing. OAB has been proposed to have a greater negative effect on the health related quality of life (HRQoL) than diabetes mellitus, hypertension, asthma or depression. Various hypotheses have explained the pathophysiology of OAB symptoms but no concrete evidence exists which supports these hypotheses. In recent years, subclinical urinary infection has been explored as an etiological factor for OAB symptoms, but robust data is still lacking. 15 The comparison of agar dilution, broth micro dilution and Etest for susceptibility testing of Bacteroides species and the detection of genes The aim of our study is to investigate whether women with OAB symptoms (and no evidence of overt clinical infection by conventional Mid Stream Urine for culture) have significant markers of subclinical infection when compared to individuals with no OAB symptoms. We aim to compare the urine samples of patients with OAB with healthy controls. We will be looking for the following markers of infection 1) microscopy of fresh urine for pus cells 2) culture of urine for bacterial growth 3) Detection of inflammatory biomarkers in the urine. 4) Molecular testing for bacterial DNA in the fourth sample. Metronidazole resistance is still considered rare in anaerobic isolates from clinical specimens; however, since January 2013 the Anaerobic Reference Unit in Cardiff has received had an increase in the number of referrals of metronidazole resistant Gram negative rods, mainly of the genus Bacteroides. As metronidazole remains the basis of treatment for anaerobic infections, the development of resistance may have an effect on 11 Mr Trefor Morris (Lead Biomedical Scientist, Anaerobe Reference Laboratory, Public Health Wales) Trefor.Morris@wales.nhs.uk conferring antibiotic resistance in these species 16 Models for Access to Maternal Smoking cessation Support: An evaluation of the effectiveness of new models of service delivery to increase engagement of pregnant smokers in stop smoking services patient management and outcome. Antimicrobial susceptibility for minimum inhibitory concentration (MIC), to show if the antibiotic will be effective or not to the bacteria of Meropenem, Clindamycin, Amoxixillin/clavulanate, Pipercillin/tazobactam and metronidazole will be conducted using agar dilution, gradient strip and Trek plates which are presently only validated for B. Fragilis to see the sensitivity of the organisms to the different antibiotics and to see if they all confer the same results. PCR will be used for detecting resistance genes an individual gene basis to see if the sensitivity patterns shown by the three different antimicrobial testing methods are as expected for the resistance genes present within the organism. There is currently no multiplex Polymerase Chain Reaction (PCR) available to screen several genes in a single transaction but with the potential to develop one. The study aims to compare the effectiveness of new models of service delivery for specialist smoking cessation services for pregnant women with existing services delivered by Stop Smoking Wales. The new models of service delivery (pilots) will be implemented over a maximum 12 month period in selected sites across four health boards in Wales using an integrated approach between maternity services, local public health teams and stop smoking Wales. Core recommendations from NICE guidance will be implemented across pilot and usual care sites within the participating Health Boards (National Institute of Health and Clinical Excellence 2010). In usual care sites, stop smoking support for pregnant women will be 12 Siobhan Jones (Consultant in Public Health/Associate Director of Public Health, Public Health Wales) siobhan.jones2@wales.nhs.uk Bennett, L, Grant, A, Jones, S, Bowley, M, Heathcote-Elliott, C, Ford, C, Jones, A, Lewis, R, Munkley, M, Owen, C, Petherick, A, and Paranjothy, S. (2014). Models for Access to Maternal Smoking cessation Support (MAMSS): a study protocol of a quasi-experiment to increase the engagement of pregnant women who smoke in NHS Stop Smoking Services. BMC Public Health 14 delivered by the existing Stop Smoking Wales service. In pilot sites, support will be provided by a maternity support worker, a midwife or a dedicated Stop Smoking Wales advisor and will be delivered more flexibly than in usual care sites. Each health board has identified pilot and usual care sites based on geographical areas or community midwife teams depending on how maternity care is organised in the Health Boards. Routine data collected by Health Board maternity teams and Stop Smoking Wales for a cohort of women who participate in either a pilot or usual care service across the four Health Board areas will be extracted, linked and analysed for a maximum 12 month period. 17 Examining the effect of antifungal treatment and prophylaxis on the detection of molecular markers of invasive The primary outcome is engagement with specialist smoking cessation services defined by having at least one face to face therapeutic contact. Secondary outcomes are pregnant smokers who set a quit date; pregnant smokers who quit at 4 weeks follow up (CO verified); smoking status of pregnant smokers during third trimester and birth outcomes (birth weight, gestational age). Process measures will assess fidelity to protocols, feasibility acceptability, maintenance and sustainability should the pilots be rolled out on a wider level. This project is led by the University Hospital of Dr Lewis White (Principal Clinical Wuerzburg in Germany with an overall aim to Scientist, Public Health Wales) determine the effect of antifungal drugs on the Lewis.White@nhs.wales.uk detection of molecular markers for invasive aspergillosis (IA) in patient samples. The major focus of this project is a clinical study of the effects of antifungal treatment on PCR-based 13 (1):pp.1041 Grant, A, Lewis, R, Jones, S, and Paranjothy, S. (2014). Who should support pregnant women to quit smoking? Early findings from a quasi-experiment building upon NICE guidance: Models for Access to Maternal Smoking cessation Support (MAMSS) (PMM58). [Poster presentation]. In:Perinatal Medicine 9 June 2014 - 11 June 2014, Harrogate. Arch Dis Child Fetal Neonatal Ed 99 (Suppl 1):pp.A142. aspergillosis (IA) with the aim of improving PCRbased detection to compensate for any deleterious effects from treatment (Polymerase chain reaction) diagnostic methods. Specifically, the project will examine the effect of antifungal drugs on the in vitro release of DNA and galactomannan (GM) which is measured to determine the magnitude of drug effects on relevant biomarker detection. A murine model of IA is used to validate the in vitro results and to allow optimisation of the detection methods. The final objective is to see if the observations from in vitro and in vivo models can be used to improve detection of fungal DNA and GM in patient plasma and whole blood samples, and this is the only involvement Cardiff will have in the study. As part of the diagnostic service provided by Public Health Wales and Cardiff University laboratory services) EDTA blood and serum are routinely collected and tested by PCR and GM ELISA. PCR is performed on plasma and GM ELISA is performed on serum. Remaining samples (serum, plasma and blood cells) are prospectively stored according to local laboratory procedures for quality assurance and performance assessment purposes. 18 Can we use polymerase chain It is proposed to test these stored blood cells and serum specimens by PCR and stored plasma by GM ELISA. The aim is to test 1000 blood samples (5ml EDTA anticoagulated blood) from 50 patients who are undergoing HSC (haematopoietic stem cell transplant) until day +100 post transplantation, over a 6 month period. All eligible specimens will be shipped to University Hospital Wuerzburg for subsequent anonymous PCR analyses. Aims: The aim of this study is to determine whether Professor Rosemary Barnes non-invasive samples such as blood, sputum or (Honorary Consultant Medical 14 reaction (PCR) on non-invasive specimens to diagnose Pneumocystis jirovecii pneumonia? nasopharyngeal swabs could be used as an alternative to invasive specimens to diagnose Pneumocytis pneumonia (PCP). Methods: For each bronchoalveolar lavage (BAL) sample that is found to be PCP PCR positive, our aim is to check for alternative sputum, serum or nasopharyngeal swabs sent to the microbiology laboratory for the same patient within the same time period (+/- 2 days) that were initially sent for alternate tests. By also performing the same PCP PCR on these non-invasive specimens and comparing this data with that obtained from the corresponding BAL, we can work out the sensitivity and specificity of PCP PCR in these non-invasive specimens. Primary outcome measure: To determine whether PCR can be used on non-invasive samples for the diagnosis of PCP pneumonia. 15 Microbiology, Public Health Wales) Rosemary.Barnes@wales.nhs.uk