Biomedical Laboratory techniques - Course A - Roma Azienda Policlinico Umberto I

Activity type

Scienze e tecniche di laboratorio biomedico

SSD

MED/46

Year

N/D

Semester

N/D

CFU

2

Hours distribution

20 classroom hours

Lecturers

SONIA CONI

Activity type

Scienze medico-chirurgiche

SSD

MED/08

Year

N/D

Semester

N/D

CFU

2

Hours distribution

20 classroom hours

Lecturers

FRANCESCA ROMANA BUTTARELLI

Activity type

Scienze e tecniche di laboratorio biomedico

SSD

MED/05

Year

N/D

Semester

N/D

CFU

2

Hours distribution

20 classroom hours

Lecturers

GIUSEPPE GIANNINI

Main teaching objectives:
The course is meant to provide the fundamental knowledge on the most relevant and most updated methods in molecular genetics applied to diagnostic, prognostic and predictive issues in human pathology.

By the end of the course, the student must:
 Know the organization of the molecular diagnostic laboratory, the main and most updated techniques applied to the diagnosis of genetic and neoplastic diseases, the instrumentation used, the procedures for the technical validation of the tests.
 Be able to evaluate their potential application to different sample types, for diagnostic, prognostic and predictive purposes.
 Own the intellectual skills to solve specific diagnostic issues being able to design specific laboratory approaches.
 Be able to critically discuss on the potential and limitations of different laboratory methods with students and professors.

A basic knowledge of biology, genetics, histology is required.

Module: TECHNICAL SCIENCES OF LABORATORY MEDICINE
Diagnostic Techniques for Autosomal Recessive Hereditary Diseases:
Phenylketonuria and Cystic Fibrosis: Laboratory Analysis and Diagnosis
Diagnostic Techniques for Sex-Linked Hereditary Diseases:
Fragile X Syndrome: Laboratory Analysis and Diagnosis
Laboratory Diagnostics of Autoimmune and Allergic Diseases
Molecular Biology Techniques and Their Diagnostic Applications:
Qualitative and Quantitative Methods for Nucleic Acids Amplification
Techniques for Protein Extraction and Analysis from Cells and Tissues: Membrane-Based and Immunofluorescence Methods
Nucleic Acid Sequencing Techniques
“Omics” Techniques: Total RNA and Single-Cell RNA Analysis



Module: MOLECULAR DIAGNOSTICS ON FABRIC
Proteomics and genetic analysis in pathological anatomy. In-situ biomarker analysis. Analytical methods on extracted DNA.
In-depth analysis of different methods of DNA extraction, purification and quantification, pro and cons.
Practical microscopy, observation and evaluation of samples using immunohistochemistry and FISH.
Gene alterations and mutations that can be analysed for diagnostic purposes on tissue in neoplasms.
Fluorescence in situ hybridisation (FISH); chromogenic in situ hybridisation (CISH); SISH.
Epigenetics and methods for analysing DNA methylation.
Pyrosequencing vs Sanger; Next Generation Sequencing in genes involved in response to therapy in neoplastic syndromes.
Tissue Micro Array (TMA). Biological banks.
Droplet Digital PCR for the identification of low-frequency point mutations in neoplasms. Identification of point mutations at the point of care (POC) using Nanopore sequencing.



Module: MOLECULAR DIAGNOSTIC METHODOLOGY
Contents:
 Brief overview of genetic pathology. Review of the most relevant methodological achievements in molecular biology/molecular genetics.
 Traditional and updated methods to detect structural and numerical chromosomal aberrations.
 Traditional and updated methods to detect DNA rearrangements.
 Traditional and updated methods to detect point mutations.
 Set-up of pathology-related mutational screenings.
 Evolution of Sequencing technologies.
 Application of mutational screening approaches to different medical issues, with a particular focus on molecular oncology.
 High throughput molecular methods in molecular oncology.

Module: TECHNICAL SCIENCES OF LABORATORY MEDICINE
Suggested books:
Molecular Methodologies» Mauro Maccarone (Zanichelli)
“Clinical biochemistry and laboratory medicine” Ciaccio - Lippi (Edises )
Scientific articles from the ncbi/Pubmed database cited in the slides



Module: MOLECULAR DIAGNOSTICS ON FABRIC
F. Amaldi, P. Benedetti, G. Pesole, P. Plevani, Tecniche e metodi per la biologia molecolare, Casa Editrice Ambrosiana. Distribuzione esclusiva Zanichelli
Terry A. Brown, Biotecnologie molecolari, ed. Zanichelli
Molecular Diagnostic, W. B. Coleman, G. J. Tsongalis, Humana Press



Module: MOLECULAR DIAGNOSTIC METHODOLOGY
Suggested textbooks:
Any book of uman Genetics, plus occasional readings on the following books:

Molecular Diagnostic
W. B. Coleman, G. J. Tsongalis
Humana Press

Introduzione alla Medicina Molecolare
Ross Dennis W.
Springer

Human Molecular Genetics
Strachan T, Read . P.

Module: TECHNICAL SCIENCES OF LABORATORY MEDICINE
N/D
Module: MOLECULAR DIAGNOSTICS ON FABRIC
N/D
Module: MOLECULAR DIAGNOSTIC METHODOLOGY
Updated bibliography will be eventually communicated during the lectures

Lectures, elective activities eventually including a laboratory experience.

Compulsory attendance

The exam consists of an oral discussion of topics covered during the lessons in order to assess the student's understanding of the course content and critical thinking skills.

Module: TECHNICAL SCIENCES OF LABORATORY MEDICINE
N/D
Module: MOLECULAR DIAGNOSTICS ON FABRIC
N/D
Module: MOLECULAR DIAGNOSTIC METHODOLOGY
N/D

Module: TECHNICAL SCIENCES OF LABORATORY MEDICINE

• DNA amplification: classical, quantitative, and digital approaches


• liquid biopsy and non-invasive prenatal tests


• DNA sequencing: First, Second and Third generation


• Protein extraction: western blot technique, immunofluorescence, E.L.I.S.A. and diagnostic applications


• Autosomal recessive hereditary diseases: Cystic fibrosis laboratory diagnosis


• Autosomal recessive hereditary diseases: phenylketonuria laboratory diagnosis


• thern blot and fragile X syndrome diagnosis


• Laboratory diagnosis of autoimmune and allergic diseases


• “Omic” techniques: transcriptomic analysis. RNA seq, Single Cell RNA, Spatial Transcriptomics

• Proteomics and genetic analysis in pathological anatomy. In-situ biomarker analysis. Analytical methods on extracted DNA.


• In-depth analysis of different methods of DNA extraction, purification and quantification, pro and cons. 


• Practical microscopy, observation and evaluation of samples using immunohistochemistry and FISH.


• Gene alterations and mutations that can be analysed for diagnostic purposes on tissue in neoplasms.


• Fluorescence in situ hybridisation (FISH); chromogenic in situ hybridisation (CISH); SISH.


• Epigenetics and methods for analysing DNA methylation.


• Pyrosequencing vs Sanger; Next Generation Sequencing in genes involved in response to therapy in neoplastic syndromes.


• Tissue Micro Array (TMA); Biological banks.


• Droplet Digital PCR for the identification of low-frequency point mutations in neoplasms. Identification of point mutations at the point of care (POC) using Nanopore sequencing.

• Introduction to molecular diagnostics


• Genetic diseases and DNA mutations


• Methods to detect numerical and structural chromosomal aberrations


• Methods to detect point mutations


• PCR, Quantitative PCR and its applications


• Molecular diagnostics for predictive oncology


• Molecular diagnostics for hereditary tumors


• Next Generation Sequencing