Syllabus
The lectures are given in Czech language only.
Practical course in virology takes place as a two-week tournament. The course is designed for students in virology and molecular biology. The capacity of one course is a maximum of 18 students and is limited by the difficulty of the methods performed. During the course, students will learn how to work with infectious biological material in a laboratory with the biosafety level 2 (BSL-2) and learn the habits necessary to work with potential human pathogens. Within the course there are 3 model practical tasks, which are conceived so as to methodically represent problems that the student can encounter in basic research and biomedical fields. It is ensured that the methodologies carried out do not overlap with the content of practical courses in molecular biology and genetics, but become an extension of these courses. The course also includes a series of short lectures on the theoretical foundations of methods and viral expression systems.
Task # 1: IDENTIFICATION OF VIRUS IN BIOLOGICAL SAMPLE. Working with tissue cultures detection of known / unknown virus and determination of productive vs. abortive infection (viral transformation); DNA and protein analysis. Methods: DNA isolation, comparison of PCR-free and rolling-circle amplification (RCA) methods in infection diagnostics; ELISA
Task # 2: QUANTIFICATION OF VIRUS. Methods: Comparison of several methods of virus quantification, plaque titration, titer determination by immunofluorescence, end-pint dilution method, observation of infection symptoms.
Task # 3: PREPARATION AND PURIFICATION OF VLPs (VIRUS-LIKE PARTICLES) AND ANALYSIS OF INTERFERENCE IN INFECTION. Procedure: Preparation of VLPs using a baculovirus expression system and analysis of murine polyomavirus infection in the presence of isolated VLPs (to determine if there is a competitive inhibition of infection). Methods: isolation and purification of VLPs (ultracentrifugation in CsCl gradient), immunodotblot or Western blot, quantification of VLPs by haemagglutination (or protein determination). Contrasting of samples by negative staining, electron microscopy, immunodetection of infected cells -immunofluorescence or flow cytometry.
Annotation
Two-week group practical classes are designed to illustrate the topics covered in the lecture and to train students with diploma specialization in virology. Students gain familiarity with the fundamental laboratory methods commonly used not only in virology (work with cell cultures, isolation, purification and quantification of viruses, analyses of viral products) and with the use of viruses in the molecular biology (baculovirus expression vectors) and biomedicine.