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Evaluation of ELISA Methods and determination of concentration of ICA, IA2 and GAD in Diabetes Mellitus Type 1
Bosnia and Herzegovina (BoHeMSA) - University of Mostar, Mostar
Laboratory Diagnostic Institute, School of Medicine University of Mostar
PhD Ivanka Mikulić, MD
Kristina Ljubić; Vinka Barać; Ana Ćuk
English, Croatian, Bosnian, Serbian
Type of Research Project
- Basic science
What is the background of the project?
Diabetes mellitus (DM) is a group of metabolic diseases that are characterized chronic hyperglycemia due to defect in insulin secretion or a defect in its operation or the existence of both of these disorders. The disease manifestation precedes the long-lasting, covert autoimmune process of selective destruction of pancreatic beta cells, the so-called pre-diabetes. It is unclear whether the destruction of beta cells is continuous or intermittent. Molecular mechanism of occurrence is insufficiently known. Excess apoptosis is believed to be the basis for the formation of DM. Antibodies to Langerhans Island Cells, so-called. ICA(islet cell antibodies) - circulating antibodies to the cytoplasm of Langerhans' islet cells, IA (insulin autoantibodies) - antibodies against endogenous insulin, GADA - antibodies to the membrane protein beta cell and IA-2 - antibodies to tyrosine phosphatase, years before the event. They are referred to as immunoassays and are the basis of claims about a long period of predial diabetes. By Paradini study the frequency of positive results in recent-onset (less than 6 months) DM type 1 patients was 80.0% for GADA, 62.9% for IA-2 and 82.9% for GADA and/or IA-2. The long-duration type 1 diabetes (more than 12 months) subjects presented frequencies of 54.1% for GADA and IA-2, and 67.5% for GADA and/or IA-2 antibodies. With ELISA we are practically able to predict onset of Diabetes Mellitus 1. In order to that serious measurements of ICA, IA2 and GADA need to be done. Goal is to evaluate is there and at which rate correlation between antibodies and Diabetes Mellitus 1.
What is the aim of the project?
To measure concentration and quantify correlation between ICA (islet cell anti)bodies), IA-2 (antibodies to tyrosine phosphatase), GADA (antibodies to membrane protein beta cell) and Diabetes Mellitus 1.
What techniques and methods are used?
ELISA – Enzyme-linked immunosorbent assay will be used to detect ICA (islet cell antiboides), IA-2 (antibodies to thyrosine phosphatase) and GADA (antibodies to membrane protein). By analyzing collected serum, which will be randomly stratified, of Diabetes Mellitus 1 patients and compared with healthy samples we shall calculate the correlation between antibodies and DM1. Main tools used during research project will be : ELISA and Microsoft Excel. Microsoft Excel is used for statistical calculations such as correlation and significance. Project has three stages. First one involves education and preparation, second involves analysis of serum and data input. Third stage is data presentation.
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
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Student will be preforming ELISA on collected serums. During this process student will not go and personally collest serum but rather have final product dilivered to the laboratory. Together with supervisor studeent will conduct experiments and record given data in Microsoft Excel. After every working day student will be required to clean and test sterility of working are as well do the calibration the next day (mentor shall introduce student how machines are calibrated). handling with basic laboratory techniques (ELISA used for measurements). On final week of exchange student will write a scientific report and present it to the supervisor in order to gain certificate.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Student will be introduced with the methods of ELISA, how to handle equipment and calibrate for the first seven days by mentor. Student will be highly prepared to work together with seniors with samples and measurements.
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 scientific report - The student’s name will be mentioned in a future publication - The student will have the opportunity to present the results together with the supervisor at a conference
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Student will acquire knowledge which will introduce them with ELISA, Diabetes Mellitus 1 and correlation with antiodies; search the literature; data analysis; data presentation; skills in writing the paper
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- A short history; principles; and types of ELISA; and our laboratory experience with peptide/protein analyses using ELISA. Aydin S. 2015 Oct;72:4-15. doi: 10.1016/j.peptides.2015.04.012. Epub 2015 Apr 20
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