110 Abstracts / International Journal of Infectious Diseases 53S (2016) 4–163 20.035 Fast-track Diagnostics Enteric fever assay: From pre-enrichment to multiplex real-time PCR E Nicolas a,∗ , S Pouzol b , L Fabre c , F.-X Weill c , H Endtz d , B Carman e a Fast-track Diagnostics, Esch-sur-Alzette/LU Fondation Mérieux, Lyon/FR c Institut Pasteur, Paris/FR d ICDDR,B, Dhaka/BD e Fast-track Diagnostics, Luxembourg/LU b Purpose: Enteric fever due to Salmonella remains a significant public health problem, predominantly in children in developing countries Typhoid fever affects more than 20 million people per year associated to about 300,000 deaths These high rates are mainly due to the lack of fast, reliable and inexpensive diagnostic assays Diagnosis of typhoid remains a challenge because a low bacterial load can be responsible of an illness and also because the volume of blood that can be collected from children is limited To overcome the problem of sensitivity, the first goal of this study was to define an appropriate pre-enrichment method of the collected sample before nucleic-acid extraction The second objective was to develop a sensitive multiplex real-time PCR assay for detection of the common etiological agents of Enteric fever: Salmonella enterica spp., Salmonella typhi and Salmonella paratyphi A Methods & Materials: To determine the best pre-enrichment medium, we spiked S paratyphi A within blood samples, added the enrichment medium, cultured for h at 37 ◦ C, performed nucleic acid extraction and determined the yield of specific nucleic acids by real-time PCR We tested several commercial enrichment broths and a home-made TSB medium supplemented with Oxgall (inhibitor of Gram+ bacteria) The results show that the Selenite broth (Copan) and the TSB-Oxgall medium are the most efficient pre-enrichment methods of free bacteria in blood samples Results: In parallel we developed a multiplex real-time PCR assay that distinguish between infection by S enterica spp., S typhi and/or S paratyphi A (+ internal control) This assay shows a limit of detection between 1E+02 and 1E+03 copies/ml and a linearity from 1E+09 to 1E+03 copies/ml for all pathogens The specificity of the assay was validated with various negative samples which did not generate any positive signals Furthermore, different positive materials containing bacteria, parasites, and viruses were evaluated with this assay and no other than the expected bacteria were detected Conclusion: In conclusion, this study shows the development of an efficient diagnostic pipeline that combines a quick preenrichment of infected blood samples with a sensitive real-time multiplex PCR assay that could be used as a reference diagnostic method for Enteric fever infectious agents http://dx.doi.org/10.1016/j.ijid.2016.11.275 20.037 Effectiveness of measuring blood antibody titer and biofilm-forming ability in Staphylococcus aureus bacteremia M Yamashita Department of Medical Engineering Faculty of Health Care Science Himeji Dokkyo University, Himeji/JP Purpose: To examine the effectiveness of measuring anti-Staphylococcus aureus antibody titers in the blood and biofilm-forming ability of the pathogen as supporting evidence for the diagnosis of bacteremia in patients with S aureus infection Methods & Materials: Anti-S aureus antibody titers in the blood were measured by ELISA using protein A-deficient S aureus The biofilm-forming ability was measured using the tissue culture plate method Results: Regarding anti-S aureus antibody titers, the patient group showed significantly higher IgG and IgA antibody titers compared with the healthy control group There was no significant difference in IgM antibody titers between the patient and healthy control groups Furthermore, significantly higher IgG and IgA antibody titers were observed in the bloodstream infection group compared with the respiratory tract infection group In the bloodstream infection group, the IgA antibody titers of the 27 patients for whom multiple measurements could be obtained over time were elevated prior to positive blood culture and subsequently decreased; of these patients, six with a poor prognosis died In patients for whom the blood culture was negative, IgA antibody titers tended to be elevated or unchanged There was no marked change in IgG antibody titers With regard to the biofilm-forming ability, there was no significant difference between MSSA and MRSA However, the examination of a possible relationship with prognosis in bacteremia cases by surveying the inpatient medical records of items, including leukocyte count, CRP, the presence or absence of fever, and outcomes, revealed that patients with a poor prognosis exhibited a significantly higher biofilm-forming ability than those with a good prognosis It is believed that an OD value of 0.8 or above indicates strong adhesiveness Furthermore, no relationship was observed between IgG antibody titer, IgA antibody titer, and biofilm-forming ability Conclusion: Patients with bacteremia show significantly higher anti-S aureus antibody titers, and it is believed that the production of large volumes of IgA antibodies can improve their condition Conversely, low titers suggest a poor prognosis Results also suggested that biofilm-forming ability is associated with prognosis in patients with bacteremia http://dx.doi.org/10.1016/j.ijid.2016.11.276 20.041 Interactive Identification Key for Female Mosquitoes (Diptera: Culicidae) of Euro-Mediterranean and Black Sea Regions F Gunay a , M Picard b,∗ , V Robert b a Hacettepe University, Faculty of Sciences, Department of Biology, Division of Ecology, Ankara/TR b IRD, MIVEGEC, Montpellier/FR Purpose: In the context of the recent introduction of invasive species, an urgent need for a user-friendly identification key for mosquitoes has resurfaced among entomologists and health care officials As the MediLabSecure medical entomology network team, our experience has shown that it is essential to have a tool that can be used freely, online and offline, by both specialists and nonspecialists Methods & Materials: 134 mosquito species are included in this tool These mosquitoes are distributed in 65 countries in Europe, North Africa, the Black Sea area and Middle East Morphological descriptors for female identification are used to build a comprehensive database This morphological database was consequently edited using the computer software Xper2 , a useful tool to create interactive and easy to use identification keys Abstracts / International Journal of Infectious Diseases 53S (2016) 4–163 Results: This interactive morphological identification key was developed using 57 morphological characters (descriptors): 11 on the head, 15 on the thorax, on wings, 22 on legs and on the abdomen Geographic distribution is another very useful character where more than one country could be selected All descriptors have detailed description, along with a picture or an illustration Priority was to line up the descriptors, considering non-experts will use the tool so characters that are more visible than others come first As an option there are three different models that can choose the question with the higher discriminant power that leads to a quicker identification It allows the user to see the descriptors, states, definitions, images and all taxa on the same page Conclusion: Training sessions organized in the frame of the MediLabSecure Project, with participants from many of the selected countries in this task, have brought to light that a tool to train inexperienced users is a must This identification key will be available in January 2017 This multi-entry key will allow using descriptors the user can observe This will help when some body parts are missing on the sample since it’s possible to select a group of descriptors Future plans are to include a key for fourth instar larva and to perform validation tests on entomologists with variable degrees of experience http://dx.doi.org/10.1016/j.ijid.2016.11.277 20.043 Implementation of GastroBusters in Toronto: Canada’s First Online Reporting System for Foodborne Illness A.A.A Arthur ∗ , S Ota, E Gournis Toronto Public Health, Communicable Disease Control, Toronto/CA Purpose: Foodborne illness (FBI) is greatly under-reported to public health Toronto Public Health implemented an online reporting option, GastroBusters, for the public in July 2015, in preparation for the Pan and Parapan American Games (PPAG) Mass events increase risk of FBI and also involve visitors who may not access local health services unless acutely ill System objectives included: (1) improved ability to detect FBI clusters/outbreaks, (2) more timely data for public health interventions, (3) better estimates of FBI in Toronto, and 4) potential use as a legacy system for routine surveillance Methods & Materials: FluidSurveys was used to create a user friendly, secure, online data collection tool Users had the option to provide their contact information, with clear indication that this would permit contact by a public health investigator A 3-day food history was requested, with prompts for large events (e.g., PPAG) that were attended Nominal reports were escalated for followup within two hours of receipt; exposure data from both nominal and non-nominal reports were analysed with other surveillance data sources to identify commons exposures Data were collated in MS Access and analyzed using SASv9.3 GastroBusters was piloted during the PPAG and then evaluated after months Results: Between August 1, 2015 and January 31, 2016, 211 food poisoning reports were submitted through GastroBusters; 79% were not reported by another surveillance data source (e.g., telephone or food premise complaint) Two-thirds (67%) provided contact information permitting public health follow-up Eightyfive reports prompted a food premise inspection, to identify and mitigate food safety infractions Of these, 15 (18%) identified critical infractions and three premises were closed No widespread community outbreaks or clusters of illness were detected by any 111 surveillance sources during the pilot period, including during the PPAG Conclusion: Providing an online reporting option yielded valuable and timely information on FBI in Toronto Integration with a food safety and premise inspection program can trigger actions needed to protect the public from further illness GastroBusters is the first online FBI reporting tool implemented in Canada, and is a valuable legacy of the PPAG that can be used to complement other surveillance data sources http://dx.doi.org/10.1016/j.ijid.2016.11.278 20.044 Harvesting Real Time and Historical Disease Outbreak Data from the ProMED-Mail Database: Pitfalls and Proposed Solutions C.E.B Carslake University of Bristol, Veterinary Science, Bristol/UK Purpose: Internet based disease reporting systems are potential sources of vast amounts of data that can be incorporated into epidemiological models investigating disease distribution and drivers However steps must firstly be taken in order to address the veracity, volume and variety of the available data This poster details our attempt to harvest and transform online disease outbreak reports from Pro-MED mail into reliable and usable data detailing infectious disease outbreaks in humans and animals at a country level Methods & Materials: ProMED-mail issues reports regarding disease outbreaks as well as requests for information, collegial notifications (conferences, publications) and other information Unfortunately for the epidemiologist the type of information contained within the report, as viewable through Health Map, is not immediately ascertainable In addition the information contained within the species field is often unreliable and the location reported is occasionally that of the information source not the outbreak We propose an automated method to harvest infectious disease outbreak information from the ProMED-mail reports Firstly individual reports are grouped into disease events (a series of individual reports of the same disease occurring within a specified time frame and location) thereby eliminating individual reports that not relate to disease outbreaks Secondly an algorithm infers the species affected by scanning the grouped reports for key words and checking the compatibility of the species identified with the disease reported Results: Grouping reports of the same disease at the country level with a maximum of 21 days between each report transforms the 65492 reports over 10 years into 9436 disease events containing 51023 reports Each disease event can be reliably attributed to an outbreak The grouping of the reports into disease events allows a more accurate representation of the species affected The original data did not identify a species in 14% of reports Our algorithm identified a species in all but 1% of disease events Conclusion: Our methodology allows the creation of a database documenting the distribution of animal and human disease outbreaks worldwide We aim to integrate this ‘One Health’ database with a wealth of other data resources in order to characterise and quantify disease drivers through data modelling techniques http://dx.doi.org/10.1016/j.ijid.2016.11.279 ... regarding disease outbreaks as well as requests for information, collegial notifications (conferences, publications) and other information Unfortunately for the epidemiologist the type of information... of reports Our algorithm identified a species in all but 1% of disease events Conclusion: Our methodology allows the creation of a database documenting the distribution of animal and human disease... harvest infectious disease outbreak information from the ProMED-mail reports Firstly individual reports are grouped into disease events (a series of individual reports of the same disease occurring