Human Cytomegalovirus Infections

Objectives

1. Knowledge of CMV infections. Application of different diagnostic techniques depending on the host and clinical context (immunocompetent, immunocompromised, pregnant, newborn);

2. Knowledge of the application of real time PCR to the diagnosis of cytomegalic infection;

3. Contact with the diagnostic methods used for CMV (Culture; Antigenemia; Molecular Biology Methods: PCR "in-house technique"; Real-time PCR; Serology: routine immunoassay assays, confirmatory tests - Western blot, IgG avidity);

4. Recognition of the importance of genotypic detection of ganciclovir resistance. Knowledge of the different steps of this detection.

General characterization

Code

10210

Credits

6.0

Responsible teacher

Paulo Paixão

Hours

Weekly - 3

Total - 38

Teaching language

Português

Prerequisites

Available soon

Bibliography

- Paulo Paixão, Sofia Almeida, Paula A. Videira, Dário Ligeiro, Teresa Marques. “Screening of congenital cytomegalovirus infection by a real time PCR in urine pools”. European Journal of Pediatrics, 2012; 171 (1): 125129.

- Kotton CN, Kumar D, Caliendo AM, Asberg A, Chou S, DanzigerIsakov L, Humar A. Updated international consensus guidelines on the management of cytomegalovirus in solidorgan transplantation. Transplantation. 2013;96(4):33360.

- Revello MG, Gerna G. Pathogenesis and prenatal diagnosis of human cytomegalovirus infection. J Clin Virol. 2004;29(2):7183.

Teaching method

Available soon

Evaluation method

Student assessment – Written examination finally multiple choice test of 20 questions (duration 30 minutes) 50%.

Oral presentation and discussion of topics by groups of two students. Themes are distributed in 1st class and the presentation of each theme has a duration of 20 minutes 50%.

Subject matter

Themes and Contents of the lecture (s):

- Introduction to the diagnosis of CMV infections. Application of different techniques depending on the host and the clinical context (immunocompetent, immunocompromised, pregnant, newborn);

- Antivirals acting on CMV. Resistance and its detection;

- Real time PCR (general concepts and specific application to CMV);

- Discussion of clinical and laboratory cases;

- Discussion of the results obtained in practical teaching;  

Themes and Contents of the practical class (s):

- Culture: observation of flasks with characteristic cytopathogenic effects; preparation of tubes for shell-vial culture, inoculation, development and observation after incubation;

- Antigenemia: separation and preparation of slides; staining and observation. PCR (in-house technique): extraction and amplification PCR: detection of amplifiers;

- Serology: routine immunoassay assays, confirmatory tests (Western blot, IgG avidity);

- PCR (in-house technique): extraction and amplification; detection of amplifiers;  

- Genotypic detection of ganciclovir resistance: extraction, amplification and observation of sequences;

- Real time PCR: extraction, amplification, detection.

Programs

Programs where the course is taught: