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Course, academic year 2025/2026
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Practical course of diagnostic methods in virology - MB140C85
Title: Praktikum z vybraných diagnostických metod ve virologii
Czech title: Praktikum z vybraných diagnostických metod ve virologii
Guaranteed by: Department of Genetics and Microbiology (31-140)
Faculty: Faculty of Science
Actual: from 2025
Semester: winter
E-Credits: 3
Examination process: winter s.:
Hours per week, examination: winter s.:0/1, C [TS]
Capacity: 8
Min. number of students: 6
4EU+: no
Virtual mobility / capacity: no
State of the course: taught
Language: Czech
Level: specialized
Note: enabled for web enrollment
Guarantor: RNDr. Jana Šmahelová
Teacher(s): RNDr. Ingrid Poláková, Ph.D.
RNDr. Martina Saláková, Ph.D.
RNDr. Jana Šmahelová
Co-requisite : {At least one of the following subjects: MB140P75, MB140P81}
Incompatibility : MB140C69E
Is incompatible with: MB140C69E
Annotation -
This practical course will focus on molecular methods of viral detection in clinical specimens including the
interpretation of results. The methods will include extraction of nucleic acids from different type of clinical material. The students will analyze the quality and quantity of nucleic acids. They will also detect the selected genes by the polymerase
chain reaction (PCR) and/or real time PCR. The specificity of amplicons will be confirmed by sequencing.
For the detection of viral specific antibodies Enzyme-Linked Immunosorbent
Assay (ELISA) and/or indirect immunofluorescence method will be used.
The course will be held in BIOCEV, Vestec from Oct 13th to Oct 17th 2025
Last update: Šmahelová Jana, RNDr. (04.09.2025)
Literature -

See MB140P75

Last update: Poláková Ingrid, RNDr., Ph.D. (03.09.2024)
Requirements to the exam

 The course will be held in BIOCEV, Vestec from Oct 13th to Oct 17th 2025

In order to obtain credits the students have to provide the protocols per each group (2 students per group). The protocols have to include interpretation of the results. 

Last update: Šmahelová Jana, RNDr. (04.09.2025)
Syllabus -

Direct methods - PCR, qRT PCR, sequencing

Indirect methods - imunofluorescence, ELISA

 

Emphasis will be on good clinical practice. The course will mimic the situation in the clinical laboratory. 

Last update: Poláková Ingrid, RNDr., Ph.D. (29.08.2022)
Learning outcomes -

Upon successful completion of the course, the student:

Task 1: HPV Detection and Genotyping (DNA Isolation, PCR, Sequencing)

  • Performs biological material sampling (skin and oral cavity swabs) under aseptic conditions and prepares samples for molecular diagnostics.
  • Applies the manual method of DNA isolation from clinical material and explains the function of individual reagents in the lysis and precipitation process.
  • Prepares and performs the PCR reaction and interprets the examination result as negative/positive.
  • Describes the principle of Sanger sequencing and prepares samples for the sequencing reaction.
  • Analyzes obtained sequencing data (chromatograms) and uses the BLAST bioinformatics tool to compare sequences with the database for the accurate determination of the Human Papillomavirus (HPV) genotype.

Task 2: HSV Serological Diagnostics (Indirect Immunofluorescence Assay - IFA)

  • Explains the principle of indirect immunofluorescence assay (IFA) and justifies the use of the method for the detection of specific antibodies against Herpes Simplex Virus (HSV) in patient sera.
  • Performs the staining of microscopic slides, including sample dilution, washing, and fixation of the slide.
  • Evaluates slides using a fluorescence microscope, identifies positive sera, determines their titer based on specific fluorescence in infected cells, and distinguishes positive findings from background or artifacts.

Task 3: BKPyV Serological Detection (ELISA Method)

  • Defines the principle of Enzyme-Linked Immunosorbent Assay (ELISA) and describes the antigen-antibody interaction in the context of detecting anti-BKPyV antibodies in patient serum or plasma.
  • Assembles and performs individual steps of the ELISA test, including dilution and pipetting of patient samples, washing, and application of the chromogenic substrate.
  • Measures the absorbance of samples and, based on the measured values, interprets the examination result as qualitatively positive or negative.

Task 4: Quantification of BKPyV Viral Load (Real-time PCR / qPCR)

  • Isolates viral DNA from blood or body fluids using spin columns and prepares the reaction mixture for amplification.
  • Explains the principle of real-time PCR (qPCR) utilizing hydrolysis probes (TaqMan) and compares this method with conventional PCR in terms of sensitivity and quantification capabilities.
  • Analyzes amplification curves and calculates the absolute viral load (copies/ml) based on the constructed standard calibration curve.
  • Evaluates and justifies the clinical significance of monitoring BK virus load in immunosuppressed patients (e.g., after kidney transplantation) for the prevention of nephropathy.
Last update: Poláková Ingrid, RNDr., Ph.D. (07.01.2026)
 
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