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Tidsserieanalys av aktiv norovirus-infektion med RT-qPCR
Kristianstad University, Faculty of Natural Science.
2019 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Time-series analysis of active norovirus-infection with RT-qPCR (English)
Abstract [sv]

Norovirus som orsakar vinterkräksjukan är en av de vanligaste vintersjukdomarna i Sverige. Sjukdomstiden varar generellt i en till tre dagar med symptomen kräkning och/eller diarré. Till den totala sjukdomsbilden världen över gällande akut gastroenterit, bidrar norovirus med 18 %. Trots att sjukdomen är mycket vanlig är kunskapen om norovirusets förfarande till stor del okänd.Syftet med studien var att göra en tidsserieanalys, även så kallad One-Step Growth analys, av koncentrationen minus-RNA i celler som infekterats med olika koncentrationer av murint norovirus (MNV). För att detektera minus-RNA användes RT-qPCR med SYBR Green. Målet var att se om startkoncentrationerna av virus vid någon tidpunkt korrelerar med mängden minus-RNA i cellerna. Efter 4 och 8 timmar fanns ett exponentiellt samband mellan den initiala viruskoncentrationen och minus-RNA-uttrycket i cellerna. Koncentrationen minus-RNA i de infekterade cellerna ökade mellan de undersökta tiderna 4, 8 och 24 timmar. Vidare visade resultaten att det krävs 4 timmar för att minus-RNA skulle vara kvantifierbar vid en högre infektionskoncentration av viruspartiklar, medan det krävs 24 timmar för den lägre infektionskoncentrationen av viruspartiklar.

Abstract [en]

Norovirus causes winter vomiting disease and is one of the commonest cause of winter illness in Sweden. The disease period generally lasts one to three days with symptoms like vomiting and/or diarrhea. To the disease burden of acute gastroenteritis worldwide, norovirus contributes with 18 %. Even though the illness is very common, the knowledge about norovirus is poor and largely unknown.The purpose of the study was to do a time series analysis, a so-called One-Step Growth analysis, of the minus-RNA concentration in cells infected with different concentrations of murine norovirus (MNV). For the detection of minus-RNA RT-qPCR was used with SYBR Green. The goal was to correlate start concentration of virus at any time with the amount of minus-RNA in the cells. At 4 and 8 hours there was an exponential connection by the initial virus concentration and minus-RNA development in the cells. The concentration of minus-RNA in the infected cells increased between 4, 8 and 24 hours. Further, the results can be interpreted as requiring 4 hours for the higher concentrations to become quantifiable, while requiring 24 hours for the lower concentrations to become quantifiable.

Place, publisher, year, edition, pages
2019. , p. 26
Keywords [en]
RT-qPCR, SYBR Green, Norovirus, MNV, One-Step Growth Analysis, Winter vomiting disease
Keywords [sv]
RT-qPCR, SYBR Green, Norovirus, MNV, One-Step Growth Analysis, Vinterkräksjukan
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:hkr:diva-20077OAI: oai:DiVA.org:hkr-20077DiVA, id: diva2:1367302
Subject / course
Biomedical Laboratory Science
Educational program
Bachelor of Science in Biomedical Laboratory Science
Supervisors
Examiners
Available from: 2019-11-05 Created: 2019-11-02 Last updated: 2019-11-05Bibliographically approved

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