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Epidemiology and genotyping of Methicillin-resistant Staphylococcus aureus with amplicon-based Nanopore-sequencing : Creating a panel of clinically relevant genes

Methicillin-resistant Staphylococcus aureus (MRSA) is a variant of the more common Methicillin-Susceptible Staphylococcus aureus (MSSA), an opportunistic pathogen a portion of the human population carries as normal bacterial flora. When an outbreak of MRSA occurs, it is often important to determine if and how these strains are related to each other. In this report two different types of epidemiological methods were combined (namely Multi-Locus Sequence Typing and staphylococcal protein A-typing), in order to reduce workload, costs and time. A panel of resistance and virulence markers was also added to gather as much information about the culture as possible in a single analysis. To test the viability of the method extracted DNA and heat-treated bacterial cultures of both MRSA and MSSA were amplified with a curated panel of primers. These products were later sequenced with Nanopore’s MinION using the Flongle flow-cell. The method showed promise and worked as intended regarding the staphylococcal protein A-typing and the panel of resistance and virulence markers. However, the Multi-Locus Sequence Typing did still require optimization in order to be used clinically. In summary the project can be viewed as a success since it succeeded in being more time, cost and work efficient than many of its predecessors, when the problems with the Multi-Locus Sequence Typing are solved.

Identiferoai:union.ndltd.org:UPSALLA1/oai:DiVA.org:uu-508920
Date January 2023
CreatorsKoivistoinen Jonsson, Max
PublisherUppsala universitet, Institutionen för medicinsk cellbiologi
Source SetsDiVA Archive at Upsalla University
LanguageEnglish
Detected LanguageEnglish
TypeStudent thesis, info:eu-repo/semantics/bachelorThesis, text
Formatapplication/pdf
Rightsinfo:eu-repo/semantics/openAccess

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