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14D2I30 - Molecular-genetic diagnostics in veterinary medicine

Course specification
Course titleMolecular-genetic diagnostics in veterinary medicine
Acronym14D2I30
AcronymDOCTORAL ACADEMIC STUDIES
Module
Type of studyPhD academic studies
Lecturer (for classes)
  1. dr Jevrosima Stevanović
  2. dr Zoran Stanimirović
Lecturer/Associate (for practice)
    Lecturer/Associate (for OTC)
      ESPB10.0Status
      ConditionEnrolled semester in which the subject is attending
      The goalThe subject aims to provide students theoretical and practical knowledge about the application of the molecular methods and procedures in the diagnosis of animal disorders of various etiologies.
      The outcomeStudent should know potentials of the application of molecular techniques in veterinary medicine and be able to select the appropriate molecular technique in diagnosis of certain disorders in animals. Student should be able to independently perform the complete procedures used in molecular diagnostics. Student should understand the principles of applied molecular techniques, significance of the obtained results and correct interpretation.
      Contents
      Contents of lecturesTheoretical instructions: Features of pro- and eukaryotic genome. The genetic basis of animal disorders. Possibilities of application of molecular methods and techniques for diagnostic purposes. Advantages and disadvantages of molecular techniques in the diagnosis of certain disorders in veterinary medicine. Recombinant DNA technology. The basic principles of extraction and amplification of nucleic acids (Polymerase Chain Reaction-PCR). Reverse Transcription Polymerase Chain Reaction (RT-PCR) and its application in detecting causative agents of viral diseases and in gene expression. Real-time PCR - principle and application in the diagnosis of animal diseases of different etiology; introduction to the software used in processing obtained results; quantification of obtained products; examination of the expression of certain genes - application in immunology, pharmacology and genotoxicology. Sequencing - principle and application in the diagnosis of hereditary diseases in animals, the identification and determination of cause of animal diseases of different etiology. HRM (High resolution Melting) technology - the principle and application in the detection of mutations and early detection of neoplastic lesions.
      Contents of exercisesIndependent research activities: Rules of behaviour and safety measures in the laboratory. Type of sample and selection of molecular technique. Storing samples - various security measures. Sample preparation and nucleic acid extraction. Preparation of the PCR mixture, nucleic acid amplification (PCR), and optimization of the PCR protocol. PCR diagnosis of inherited disorders through detection of the alteration in the corresponding genes; and some bacterial and parasitic diseases in animals through detection of the pathogen DNA. RT-PCR diagnostics of viral diseases in animals by detecting pathogen RNA. Electrophoresis and analysis of results. Real-time PCR and optimizing computer - marking samples and control of reactions. Processing Real-time PCR results - determination of reaction efficacy, threshold and the amount of the obtained product. Absolute and relative quantification. Preparation for sequencing, performing the sequencing technique and analysis of obtained results. Application of HRM analysis. Contamination - the prevention, consequences and solving.
      Literature
      1. Đelic N, Stanimirović Z: Principles of genetics, Faculty of Veterinary Medicine, University of Belgrade, Elit-Medica, Belgrade, 2004.
      2. Kulić M, Stanimirović Z, Đelic N, Novakovic M: Human genetics. Faculty of Medicine in Foca, University of East Sarajevo, Foca, 2010.
      3. Stanimirović Z, Stevanović J: Applications of molecular-genetic analyses in veterinary medicine. Proceedings of the XXXIII Seminar for inovation of knowledge of veterinarians, Feb 24th, pp 17-33, Belgrade, Serbia, 2012.
      4. Stanimirović Z, Simeunović P, Vučićević M, Davitkov D, Stanišić Lj, Maletić M, Gajić B, Glavinić U, Stevanović J: Application of molecular genetic analyses in forensics and diagnosis in domestic and wild animals. Proceedings of the XXXVI Seminar for inovation of knowledge of veterinarians, Feb 20th, pp 137-146, Belgrade, Serbia, 2014.
      5. Pestana EA, Belak S, Diallo A, Crowther JR, Viljoen GJ: Early, rapid and sensitive veterinary molecular diagnostics-real time PCR applications. Springer, London 2010. (Original title)
      6. Primrose SB, Twyman R: Principles of gene manipulation and genomics. Wiley-Blackwell, 2009. (Original title)
      Number of hours per week during the semester/trimester/year
      LecturesExercisesOTCStudy and ResearchOther classes
      4
      Methods of teachingBlock lectures with student's individual work with mentors. Active and specific work on issues related to the student's doctoral thesis. Independent research activities: performing experiment, analysis and discussion of the results. Practical work will be performed on the existing equipment in the Laboratory for animal genetics at the Department of Biology, which fully supports the intended curriculum. A rotation system of PhD students: every other week a new tutor for a new diagnostic technique will be available. In this way, each candidate after completing the course is trained to independently perform every learned technique.
      Knowledge score (maximum points 100)
      Pre obligationsPointsFinal examPoints
      Activites during lectures10Test paper70
      Practical lessonsOral examination
      Projects
      Colloquia
      Seminars20