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Bestämning och jämförelse av lägsta detektionsintervall för odling och qPCR vid analys av Staphylococcus aureus
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.
2019 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Determining and comparing the lowest range of detection for cultivation and qPCR when analyzing Staphylococcus aureus (English)
Abstract [sv]

Vårdrelaterade infektioner (VRI) är ett ökande problem inom hälso- och sjukvården. På neonatalavdelningen på Länssjukhuset Ryhov i Jönköping har det förekommit inkonsekventa odlingsresultat vid misstänkt VRI orsakad av Staphylococcus aureus. Att förebygga VRI samt värna om patientsäkerheten kräver känsliga och pålitliga laboratorieanalyser. Syftet med studien var att bestämma och jämföra lägsta detektionsintervall för metoderna odling och qPCR med och utan anrikning i MAMSA-buljong, av S. aureus. Seriespädningar av S. aureus tillreddes och koncentrationer för ursprungsrören uträknades via viable count (VC). Odling på blodagar samt qPCR med och utan anrikning i MAMSA-buljong utfördes. Resultaten användes för att bestämma ett lägsta detektionsintervall. Odling gav ett lägsta detektionsintervall mellan 0,5–62 CFU/ml, och qPCR mellan 6400–140 000 CFU/ml. Anrikning med MAMSA-buljong innan qPCR-analys gav ett lägsta detektionsintervall mellan 0,6–140 CFU/ml. För detektion via odling räcker enstaka till tiotals CFU/ml i analysprovet. Vid qPCR behövs tusentals till hundratusentals CFU/ml i analysprovet, men vid qPCR med MAMSA-anrikning kan detektionsintervallet sänkas till nivåer jämförbara med odling. För att kunna applicera studiens resultat på den laborativa verksamheten krävs vidare studier med fler bakteriearter och replikat.

Abstract [en]

Hospital-acquired infections (HAI) are an increasing problem in health care facilities. In the neonatal intensive care unit at Länssjukhuset Ryhov in Jönköping, inconsistent cultivation results have been observed for suspected HAI caused by Staphylococcus aureus. To prevent HAI and to maintain patient safety, sensitive and reliable laboratory tests are essential. The aim of the study was to determine and compare the lowest range of detection for cultivation and qPCR with and without enrichment in MAMSA broth, for S. aureus. Serial dilutions of S. aureus were made, and the original concentration was determined through viable count. Cultivation on blood agar and qPCR with and without enrichment in MAMSA broth was performed. The results were used to determine a minimum detection range. For cultivation, a minimum detection range of 0,5–62 CFU/ml was determined and for qPCR a range of 6400–140 000 CFU/ml. For qPCR after enrichment in MAMSA broth, a lowest detection range of 0,6–140 CFU/ml was determined. For detection with cultivation single to tenths of CFU/ml were needed in the sample. For qPCR thousands to tenths of thousands of CFU/ml were needed. For qPCR with enrichment in MAMSA broth the detection range could be lowered to levels comparable with cultivation. For clinical application, further studies are needed with more bacterial species and replicates.

Place, publisher, year, edition, pages
2019. , p. 19
Keywords [en]
Sensitivity, CFU, viable count, E-swab, MAMSA
Keywords [sv]
Känslighet, CFU, viable count, E-swab, MAMSA
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:hj:diva-44576ISRN: JU-HHJ-BLA-1-20190081OAI: oai:DiVA.org:hj-44576DiVA, id: diva2:1325879
External cooperation
Region Jönköpings Län
Subject / course
HHJ, Biomedical Laboratory Science
Supervisors
Examiners
Available from: 2019-06-26 Created: 2019-06-17 Last updated: 2019-06-26Bibliographically approved

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