Learning objectives
Acquisition of knowledge on principles and practice of the main methods for the laboratory diagnosis of infections due to bacteria, fungi, parasites and viruses of medical interest.
Prerequisites
Getting through examinations of microbiology for the first and the second years of the course.
Course unit content
Bacteriology.
Methods for microscopic examination, cultural examination, identification, antibacterial susceptibility testing, antigen detection (agglutination test, immunochromatographic assay, enzyme-linked immunosorbent assay, immunofluorescence test), nucleic acid detection (in situ hybridization, polymerase chain reaction, nested-PCR, real-time PCR, RFLP-PCR), antibody detection (immunoenzymatic assay - EIA; ELFA, western blotting, flocculation and agglutination tests, immunofluorescence test, immunodiffusion assay) applied to the diagnosis of infections due to:
• Anaerobic bacteria (Clostridium, Bacteroides)
• Spirochetes (Borrelia, Brachyspira, Leptospira, Treponema)
• Gram-negative rods (Enterobacteriaceae, Vibrionaceae, Campylobacter, Helicobacter, Haemophilus, Bordetella, Legionella, Pseudomonas)
• Gram-negative cocci (Brucella, Neisseria)
• Gram-positive rods
• Gram-positive cocci
• Mycobacteria
• Chlamydiae, Rickettsiae, Mycoplasma
Mycology
Laboratory diagnosis of mycoses
• Microscopic examination
• Cultural examination
Procedures for identification of yeasts
Procedures for identification of moulds
• Serological procedures
• Methods for antifungal susceptibility testing of fungi
• Molecular biology approaches in medical mycology
Parasitology
Techniques to manipulate biological samples for the parasitological examination
Faecal samples
• techniques for the microscopic examination (wet mount, concentration techniques, permanent staining – trichromic staining and modified Ziehl-Neelsen staining
• techniques for in vitro cultivation of intestinal protozoa
• techniques for the culture-concentration procedure for larvae of intestinal nematoda
• techniques for the detection of antigens of intestinal protozoa
• techniques for the detection of nucleic acids of intestinal protozoa
Blood samples
• techniques for the microscopic examination (wet mount, concentration techniques, permanent and non permanent staining, Giemsa staining) for protozoa and elmints infecting blood and tissues
• techniques for the detection of antigens of protozoa and elmints (microfilariae) infecting blood and tissues
• techniques for the detection of nucleic acids of protozoa infecting blood
Urine samples
• techniques for the microscopic examination (wet mount, concentration techniques)
• techniques for in vitro cultivation of protozoa infecting the urinary and genital tract
serum samples
• techniques to detect specific antibodies to parasites by immunoassay, haemoagglutination assay, western blot assay.
Virology
Methods for electron microscopy, cultural examination (rapid and conventional), identification of cytopathogenic agents, antigen detection (agglutination test, immunochromatographic assay, enzyme-linked immunosorbent assay, immunofluorescence test), nucleic acid detection (in situ hybridization, polymerase chain reaction, nested-PCR, real-time PCR), antibody detection (immunoenzymatic assay - EIA; ELFA, emoagglutination test, immunofluorescence test, complement fixation, western blotting, immunoblot) applied to the laboratory diagnosis of:
• viral enteric tract infections (due to rotavirus, adenovirus, calicivirus, enterovirus)
• viral respiratory tract infections (due to influenza virus, parainfluenza, respiratory syncitial virus, adenovirus, cytomegalovirus)
• viral central nervous system infections (due to herpes viruses, enterovirus)
• viral genitourinary tract infections (due to adenovirus, herpes simplex, cytomegalovirus, papillomavirus)
• viral skin infections (due to herpes simplex virus, varicella zoster virus)
• viral systemic infections (due to cytomegalovirus, Epstein-Barr virus and human immunideficiency virus)
• viral infections of the eye (due to adenovirus, herpes simplex virus and varicella zoster virus)
• viral hepatitis (due to hepatitis A virus, hepatitis B virus and hepatitis C virus)
• congenital and perinatal viral infections (due to rubella virus, cytomegalovirus, herpes simplex virus, human immunideficiency virus)
Bibliography
M. Bendinelli, C. Chezzi, G. Dettori, N. Manca, G. Morace, L. Polonelli, M.A. Tufano.
Microbiologia Medica. Monduzzi Editore
A. Balows, W.J. Hausler Jr., E.H. Lennette. Laboratory diagnosis of infectious diseases - Principles and Practice. Springer-Verlag
Garcia e Bruckner. Diagnostic medical parasitology. ASM ed.
Diagnostic procedures listed in the protocols used during practical skills course.
Other information
Teachers: Prof. Maria Cristina Medici; Prof. Adriana Calderaro; Prof. Stefania Conti
