MOLECULAR DIAGNOSTICS

Degree course: 
Corso di Second cycle degree in Biotechnology for the Bio-based and Health Industry
Academic year when starting the degree: 
2022/2023
Year: 
2
Academic year in which the course will be held: 
2023/2024
Language: 
English
Credits: 
6
Period: 
First Semester
Standard lectures hours: 
48
Requirements: 

Knowledge of genetics and molecular biology methods (particularly PCR) and basic knowledge of microbiology and immunology are required.

Final Examination: 
Orale

The learning outcomes will be assessed through a single oral exam of approximately 30 minutes. A presentation of specific articles (not mandatory but contributing to the final mark) can also be agreed upon with the students. The oral exam will consist of open questions on the topics of the course, aimed at testing:

1. knowledge and comprehension of the theoretical bases underlying specific diagnostic tests, their limits, advantages, and disadvantages, when applied to specific diagnostic queries; knowledge of information to be provided in a diagnostic report for a specific molecular test;
2. ability to acquire relevant information from a scientific report and to identify critical aspects in a diagnostic method when applied to a specific clinical query;
3. ability to apply the acquired expertise to design new molecular tests;
4. knowledge of the criteria needed to set up an accredited diagnostic laboratory;
The final mark, expressed in thirties, will depend on the marks of the presentation, if foreseen (up to 2 points) and the oral exam (up to 28 points). Honors will be given only to students who demonstrate the ability to integrate the different molecular mechanisms and experimental approaches. The exam will be considered passed with a final mark equal to or higher than 18/30.

Assessment: 
Voto Finale

The main objective of the “Molecular Diagnostic Techniques” course is to make the student familiar with the procedures used in a Laboratory of Molecular Diagnostics.
The course will deal with evaluating the features of a diagnostic test (sensitivity, specificity, predictivity), with the requirements to set up a molecular diagnostics laboratory, and with the choice of the test menu to be offered. The course will also provide basic information on obtaining certification and accreditation. It will describe the techniques commonly used in diagnostics and molecular pathology laboratories, the underlying principles, advantages and limitations, and possible applications, making several examples.

At the end of the course, the student will:
- acquire knowledge on how to set up a diagnostic laboratory, to design new diagnostic tests, how to report diagnostic test results, and how to ensure quality control in diagnostics, including accreditation and certification;
- acquire knowledge of Molecular Biology and Molecular Genetics methods applied in Molecular Diagnostic laboratories, including their theoretical bases;
- read critically guidelines and papers dealing with methods or clinical applications and understand the correct execution and interpretation of a molecular test;
- ability to apply the knowledge acquired in the field of Molecular Diagnostics to make decisions on the molecular assay to be applied to particular diagnostic queries

- apply an analytical approach in the Molecular Diagnostic field;
- apply a problem-solving approach for specific Molecular Diagnostic requests

The course is organized in-class lectures and additional activities, such as visits at Molecular Diagnostics centers with accreditation, or seminars held by professionals employed in relevant Molecular Diagnostics structures, for a total of 48 hours (6 CFUs).
Class lectures will deal with the following topics:
- procedures to set up an accredited and certified laboratory of Molecular Diagnostics;
- Quality management;
- development of molecular tests, including their requirements in terms of specificity, sensitivity, and other clinical parameters;
- data to be included in the clinical report regarding molecular diagnoses;
- ethical concerns on genetic testing;
- Genome organization, types of DNA variation, and their effects;
- DNA methylation: the profile of genomic DNA methylation under physiological conditions; the enzymes involved in writing, maintaining, and erasing DNA methylation; the role of DNA methylation;
- Molecular techniques applied to molecular diagnostics as well as their main applications:
1) Basic techniques such as amplification by Polymerase Chain Reaction (PCR), including primer design; DNA ligation and nucleic acids hybridization techniques, necessary to understand more sophisticated and specific protocols;
2) Mutation detection techniques;
3) Mutation screening techniques and sequencing (including pyrosequencing and Next generation sequencing);
4) Genotyping techniques and applications (forensic analysis, transplants, pharmacogenetics);
5) Qualitative and quantitative techniques for gene expression analyses, and microbial/viral detection and identification, in order to provide information for the diagnosis, prognosis, therapy, and monitoring of therapy efficacy (including real-time PCR, ddPCR, Next Generation Sequencing, Nanostring, branched DNA);
6) Molecular cytogenetics applied to the detection of cryptic chromosomal aberrations and prenatal screening (NIPT);
7) Techniques for analysis of DNA methylation defects at gene-specific and genomic levels (tumors, imprinting defects).

The course will be enriched with several application examples. It will also briefly describe pathologies or potential etiological agents where the specific techniques can be applied for diagnostic/prognostic purposes.
Students will be engaged in an interactive discussion on selected “case reports” in which, after providing the information needed to define the clinical setting, the competencies acquired during the course will be used to choose the techniques best suited to answer the diagnostic query, taking into account limits, advantages and disadvantages of each method and the data that will be necessary to include in the diagnostic report. Finally, students will be able to demonstrate their ability to critically read an article or guidelines from the scientific literature regarding new diagnostic workflows/methods/tools and present the deriving data in class.

The course is organized in-class lectures and additional activities, such as visits at Molecular Diagnostics centers with accreditation, or seminars held by professionals employed in relevant Molecular Diagnostics structures, for a total of 48 hours (6 CFUs).
Class lectures will deal with the following topics:
- procedures to set up an accredited and certified laboratory of Molecular Diagnostics;
- Quality management;
- development of molecular tests, including their requirements in terms of specificity, sensitivity, and other clinical parameters;
- data to be included in the clinical report regarding molecular diagnoses;
- ethical concerns on genetic testing;
- Genome organization, types of DNA variation, and their effects;
- DNA methylation: the profile of genomic DNA methylation under physiological conditions; the enzymes involved in writing, maintaining, and erasing DNA methylation; the role of DNA methylation;
- Molecular techniques applied to molecular diagnostics as well as their main applications:
1) Basic techniques such as amplification by Polymerase Chain Reaction (PCR), including primer design; DNA ligation and nucleic acids hybridization techniques, necessary to understand more sophisticated and specific protocols;
2) Mutation detection techniques;
3) Mutation screening techniques and sequencing (including pyrosequencing and Next generation sequencing);
4) Genotyping techniques and applications (forensic analysis, transplants, pharmacogenetics);
5) Qualitative and quantitative techniques for gene expression analyses and microbial/viral detection and identification in order to provide information for the diagnosis, prognosis, therapy, and monitoring of therapy efficacy (including real-time PCR, ddPCR, Next Generation Sequencing, Nanostring, branched DNA);
6) Molecular cytogenetics applied to the detection of cryptic chromosomal aberrations and prenatal screening (NIPT);
7) Techniques for analysis of DNA methylation defects at gene-specific and genomic levels (tumors, imprinting defects).

The course will be enriched with several examples of application. It will also briefly describe pathologies or potential etiological agents where the specific techniques can be applied for diagnostic/prognostic purposes.
Students will be engaged in an interactive discussion on selected “case reports” in which, after providing the information needed to define the clinical setting, the competencies acquired during the course will be used to choose the techniques best suited to answer the diagnostic query, taking into account limits, advantages and disadvantages of each method and the data that will be necessary to include in the diagnostic report. Finally, students will be able to demonstrate their ability to critically read an article or guidelines from the scientific literature regarding new diagnostic workflows/methods/tools and present the deriving data in class.

Convenzionale

The course will consist of lectures. Lectures will be based on PowerPoint presentations and occasionally on movies. During the course, Seminars and a diagnostic laboratory visit will also be organized. Students will be actively engaged in activities in the last part of the course.

The lecturers are available for explanations upon appointment, to be requested by email, using student’s institutional email address.
c.kilstrup-nielsen@uninsubria.it
paola.campomenosi@uninsubria.it