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Exploratory study of melioidosis in hospitals of the social health insurance of the cities of potential high epidemiological risk for B. pseudomallei such as Piura and Tumbes.
Universidad Nacional de Piura
CI-Emerge, Center of Emerging Diseases and Climate Change Research. Universidad Nacional de Piura, Piura, Peru. Department of Public Health, Faculty of Health Sciences, National University of Piura, Urb. Miraflores S/N, Piura, Peru. 20002
Carlos Culquichicon Sanchez
Carlos Culquichicon Sanchez
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Melioidosis is an infection caused by Burkholderia pseudomallei, a bacterium widely distributed in Southeast Asia, northern Australia, South Asia (including India), China and in areas outside the classic endemic regions, most often acquired by visitors to endemic areas. It is found in soil and fresh surface waters in endemic regions. Currently, despite improved recognition and surveillance of the infection, the full extent of the global distribution of B. pseudomallei remains unclear, especially the extent to which melioidosis has spread beyond historical sites, timing and modes of global spread. Further studies are needed to understand the distribution and burden of melioidosis in high-risk clinical settings such as diabetes, chronic renal failure and chronic respiratory diseases and in its ecological niche such as moist soils, disturbed rice fields and animals interacting as livestock.
What is the aim of the project?
To describe the distribution and burden of melioidosis and other select pathogens in high-risk clinical settings and in areas with high likelihood of endemicity, through an exploratory study in hospitals of the social health insurance of the cities of potential high epidemiological risk for B. pseudomallei such as Piura and Tumbes.
What techniques and methods are used?
1. Surveillance information will be collected on those patients who meet the case definition (persons infected with Burkholderia pseudomallei) in Piura and Tumbes. 2. The enrolled participants will have their blood samples taken for identification with indirect hemagglutination test (in vitro diagnostic technique which consists of provoking the agglutination of particles artificially coated with antigens, through the reaction of anti-Burkholderia pseudomallei antibodies) and in those with positive identification, PCR and sequencing of B. pseudomallei will be performed, for the detection and quantification of the fluorescence of a reporter molecule (probe), measured along the cycle. 3. For the isolation of B. pseudomallei from soil 3.1. Checking sampling points per site: fixed interval sampling grid 3.2. Check distance between sampling points: 2.5 to 5 metres 3.3. Check soil sampling depth: 30 cm 3.4. Check weight of collected soil sample: 10 grams 3.5. Check storage during transport to the laboratory: Ambient temperature out of direct sunlight 4. For the isolation of B. pseudomallei in water: 4.1. 1 litre of water collected per sample 4.2. 10 samples per location 4.3. Use of sampling pole when necessary and plastic bags 4.4. Filtration (0.2μm pore size) 4.5.Filters used for cultivation work 5. For the detection of B. pseudomallei from soil and water 5.1. Cultivation is the standard method used with PCR as an additional option. 5.2. Use of selective media to enrich Bp (TBSS-C50 or Ashdown broth) 5.2.1. Add 10g of soil (or water filter) to 10ml Ashdown's broth 5.2.2. Mix soil samples for 30 seconds 5.2.3. Subcultivation on Ashdown agar 5.2.4. Check plate daily for Bp in suspected colonies. 6. To evaluate asymptomatic exposure we will collect blood samples from farm workers and cattle to detect anti-B. pseudomallei antibodies (IgM and IgG) by Indirect Hemagglutination Assay (IHA). 7. Human sampling: samples of elbow flexure veins will be taken keeping adequate aseptic and antiseptic measures. The samples will be obtained by trained health personnel. The biological waste will be transferred in containers of biosecurity to the laboratory of investigation in emergent diseases of the National University of Piura for its disposal. 8. Sampling in animals: samples will be taken from the external jugular vein, conserving adequate aseptic and antiseptic measures. The samples will be obtained by trained animal health personnel using a brete or rope. The biological waste will be transferred in biosecurity containers to the research laboratory for emerging diseases of the National University of Piura for disposal. 9. to examine antigens in a variety of body fluids will be used: 9.1. Latex agglutination test which consists of mixing the sample with latex drops covered with anti-Burkholderia pseudomallei antibody. 9.2. Lateral flow immunoassay (LFI), which is a diagnostic immunoassay based on the detection of the capsular polysaccharide (CPS) of B. pseudomallei within a panel of serum and urine samples from patients with melioidosis.
What is the role of the student?
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Literature survey for a project that may have taken up. Experimentation in the laboratory. Writing a paper or report. Field epidemiological operations or logistic implementation. Describe the prevalences of identified cases of melioidosis and their clinical and environmental risk factors. Experimentation in the lab. Antigen detection through a latex agglutination test. Antigen detection through lateral flow immunoassay or LFI (Diagnostic Immunoassay based on the detection of the capsular polysaccharide (CPS) of B. pseudomallei within a panel of serum and urine samples from patients with melioidosis). Execution of ELISA tests (enzyme-linked immunosorbent assay) for the detection of antibodies in human serum with Euroimmun ELISA. Writing a paper or report. Epidemiological field operations or logistical implementation. Thermal cycler handling for conventional and real time PCR (Polymerase Chain Reaction) runs.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Preliminary readings related to the research project given on the first day. Training for epidemiological field logistics. Training in report writing. Laboratory supervision. Monitoring for review of progress and/or doubts. These activities will be provided by the tutor and the scientific advisors of the local committee. we would like to have a videocall interview with the student before his/her exchange.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Just a lot of willingness to work and learn how to develop research skills. Basic handling of a thermal cycler. Management in the realization of ELISA tests. Basic knowledge in the use of statistical programs such as stata or R. Subjects passed: Epidemiology, Biostatistics,
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- 1. Currie BJ; Ward L; Cheng AC. The Epidemiology and Clinical Spectrum of Melioidosis: 540 Cases from the 20 Year Darwin Prospective Study. PLoS Negl Trop Dis. 2010;4(11):e900
- 2. Currie BJ; Dance DAB; Cheng AC. The global distribution of Burkholderia pseudomallei and melioidosis: an update. Transactions of The Royal Society of Tropical Medicine and Hygiene. 2008;102(Supplement_1):S1-S4
- 3. Inglis TJ; Rolim DB; Sousa Ade Q. Melioidosis in the Americas. The American journal of tropical medicine and hygiene. 2006;75(5):947-54.
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