Next generation sequencing data analysis in microbiology – Only Literature review part

This time I would like an offer only for the theoretical part (NOT EXPERIMENTAL).


For your greater convenience, I am attaching an outline:


1. Historical Evolution of DNA Sequencing Technologies
– Brief introduction to DNA sequencing, starting from the first generation of sequencing (Sanger method) to the new generations of sequencing.
– Review of the evolution of technologies: from Sanger technology, improvements with automated sequencing systems, and how the development of NGS has accelerated genetic research.
– Milestones in NGS technologies: From Roche 454, Illumina and SOLiD technology to newer technologies such as PacBio and Oxford Nanopore.
– Importance of NGS: Why new generation technologies have revolutionized microbiology, offering high speed, low cost and increased accuracy.


2. Conventional Microbial Identification Methods and Advantages of NGS
– Conventional methods such as cultures, bacteriological tests, PCR techniques, ELISA, and the spectroscopic MALDI-TOF.
– Advantages of NGS over conventional methods:
– Speed ​​and massive analysis
– Ability to analyze non-cultivable organisms
– Identification of new mutations and resistance genes
– Greater sensitivity and accuracy.


3. Illumina and Other NGS Technologies
– Introduction to Illumina technologies: how they work, why they are popular.
– Illumina technology details: principle of the method and detailed explanation of how it works.
– Other NGS technologies: Brief reference to other important technologies such as PacBio and Oxford Nanopore and their differences with Illumina.


4. Introduction to Staphylococcus aureus
– Importance of S. aureus as a pathogen: explanation of its role in human health, wide range of infections it causes (skin infections, pneumonia, sepsis, toxic shock).
– Antibiotic resistance: Focus on resistant S. aureus (MRSA – Methicillin-resistant S. aureus), which is one of the most dangerous and widespread multi-resistant bacteria.
– Reference to common antibiotics to which it has developed resistance, such as methicillin, vancomycin, and new resistant forms that are developing.
– Epidemiological data: How the global increase in resistance affects public health. mainly for the Netherlands (the Netherlands) – because my experimental piece will have such data, MRSA strains from the Netherlands –
-Impacts of antimicrobial resistance on public health and costs
health care


5. Basic Bacterial Genome Terminology
– Contigs: What they are and how they are derived from sequencing.
– Scaffolds: Connection and integration of contigs for genome reconstruction.
– Coverage and depth of sequencing (Depth of Sequencing): How they affect the quality of the data and the accuracy of the analysis.


6. Galaxy: Online Bioinformatics Platform
– Brief history of the Galaxy platform: How it was created and evolved as an open source platform for biological data analysis.
– Tools used to analyze the genome:
– FastQC: For quality control of sequencing data.
– FastP: For data trimming.
– Assembly (eg SPAdes): Reconstruction of the genome.
– Identification of resistance genes (ARGs) and virulence genes (e.g. through tools such as ResFinder, VirulenceFinder
7. Recent Studies Related to the Analysis of the Genome of Staphylococcus aureus:
– Studies using Galaxy to analyze the genome of S. aureus : Reference to recent research that has used Galaxy to analyze the resistance and infectivity of the bacterium.
– Important MRSA resistance studies: What we have learned so far from genetic analyzes and how data analysis is helping to understand the evolution of resistance.


8. Conclusions and Perspectives
– Reference to the importance of NGS analysis in the fight against resistant pathogens.
– Future challenges and possible use of new sequencing tools.
– The contribution of the Galaxy platform to the widespread dissemination and analysis of bioinformatics big data for microbial genetics.

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