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Jämförelse av kommersiella och InHouse kontroller för realtids-PCR vid diagnostik av Herpes simplexvirus 1 och 2
Jönköping University, School of Health and Welfare, HHJ, Dep. of Natural Science and Biomedicine.
Jönköping University, School of Health and Welfare, HHJ, Dep. of Natural Science and Biomedicine.
2018 (Swedish)Independent thesis Basic level (degree of Bachelor), 180 HE creditsStudent thesisAlternative title
Comparison of commercial and InHouse controls for real-time PCR in the diagnosis of herpes simplex viruses 1 and 2 (English)
Abstract [sv]

Herpes simplexvirus 1 och 2 orsakar godartade sjukdomar, men kan även orsaka mortalitet. Diagnostik av herpes simplexvirus 1 och 2 utförs numera framför allt med realtids- Polymerase chain reaction (PCR). I metoden amplifieras specifika deoxiribonukleinsyra(DNA)-sekvenser till miljontals kopior vilka sedan detekteras med fluorescein. I realtids-PCR sätts positiva och negativa kontroller. De positiva kontrollerna kan vara InHouse eller kommersiella. Tolkningen av resultatet inkluderar inspektion av kontrollerna. DNA utsätts för nedbrytningsprocesser av olika slag, och kan förvaras på olika sätt för att upprätthålla stabilitet. Syftet med studien var att jämföra laboratoriets InHouse-kontroller med två kommersiella kontroller, för att utvärdera vilken av dessa som var mest stabila över tid. Utvärderingen utfördes genom att analysera tre kontroller med realtids-PCR, efter att de hade förvarats i -20° C, i 5° C och i 20° C, och var spädda i TE-buffert eller i PCR-vatten. De kommersiella och InHouse-kontrollerna visade sig vara jämbördiga. Vidare studier som görs under längre tid, i större omfattning och där koncentrationerna är samma för varje kontroll, föreslås.

Abstract [en]

Herpes simplex viruses 1 and 2 which usually cause benign diseases but can even cause mortality. The diagnostics of herpes simplex virus 1 and 2 are performed with real-time Polymerase chain reaction (PCR). In the real-time PCR method, specific deoxyribonucleic acid (DNA) sequences are amplified into millions of copies which are then detected with fluorescein. Positive and negative controls are used in real-time PCR. The positive controls can be InHouse or commercial. The interpretation of the results includes inspection of the controls. DNA is subject to degradation processes of different kinds and can be stored in different ways to maintain stability. The purpose of the study was to compare the laboratory's InHouse controls with two commercial controls, to evaluate which of these were more stable over time. The evaluation was performed by analyzing the three controls with real-time PCR after they were stored in temperatures at -20° C, at 5° C and at 20° C, and were diluted in TE-buffer or in water. The commercial and InHouse controls proved to be equitable. Further studies carried out for a longer period of time, to a greater extent and where concentrations are the same for each control are suggested.  

Place, publisher, year, edition, pages
2018. , p. 31
Keywords [en]
DNA, storage, stability
Keywords [sv]
DNA, förvaring, stabilitet
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:hj:diva-40822ISRN: JU-HHJ-BLA-1-20180074OAI: oai:DiVA.org:hj-40822DiVA, id: diva2:1225316
External cooperation
Laboratoriemedicin, Länsjukhuset Ryhov
Subject / course
HHJ, Biomedical Laboratory Science
Presentation
2018-06-05, Laboratioriemedicin, pl.4, Kolven, Sjukhusgatan, 553 05 JÖNKÖPING, 10:50 (Swedish)
Supervisors
Examiners
Available from: 2018-06-27 Created: 2018-06-26 Last updated: 2018-06-27Bibliographically approved

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