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Bacteriological diagnostics of intestinal pathogens with real-time PCR - An evaluation of two molecular biology applications
Malmö University, Faculty of Health and Society (HS).
2018 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
Abstract [sv]

Tarminfektioner besitter en framträdande problematik inom primärvården, främst i form av ett ökande antal kliniska prover orsakade av dessa infektiösa tarmsjukdomar. Därav kan ett införande av molekylärbiologiska metoder vara nödvändigt som komplimenterande screeningmetod, med intentionen att förkorta befintlig analystid utan en reducering eller begränsning av analytisk sensitivitet och specificitet. Studiens utformning involverar en utvärdering av de två kommersiella realtids-PCR applikationerna: Amplidiag® Bacterial GE Assay och BD-MaxTM Enteric Bacterial Panel Assay på kliniskt retrospektiva patientprover (n = 242). Båda metoderna detekterar totalt åtta bakteriella patogener vardera, varav fem kunde mätas gentemot referensmetod. Metoderna erhöll hög specificitet på >98% för Salmonella, Campylobacter, Shigella, EHEC samt Yersinia. Och påvisade en sensitivitet på >90% för Campylobacter, Yersinia och EHEC, medan Salmonella och Shigella utfick en avvikande sensitivitet på <90%. Någon statistisk signifikant skillnad kunde inte påvisas för Amplidiag® (p-värde = 1,00) eller för BD-MaxTM (p-värde = 0,655) vid jämförelse med befintlig odlings-diagnostik. Studien visar starka argument att föreslå multiplexa realtids-PCR-analyser som grundval för en reformering av diagnostiken för tarmpatogener, då en framstående förbättring av faktorer såsom, tidsaspekt och analysmetodens prestanda erhållits.

Abstract [en]

Intestinal infections possesses a prominent problem in medical health care, primarily in the form of the increasing number of clinical patient samples caused by these infectious intestinal diseases. Hence, could a introduction of molecular biology methods may be vital as a complimenting screening method, with the intention of shortening existing analytic time without a reduction or limitation of analytical sensitivity and specificity. The study´s design involves an evaluation of the two commercial real-time PCR applications: Amplidiag® Bacterial GE Assay and BD-MaxTM Enteric Bacterial Panel Assay with the use of clinical retrospective patient samples (n = 242). Both of these methods detected a total of eight bacterial pathogens each, five of which were measured against the reference method. The methods received high specificity of >98% for Salmonella, Campylobacter, Shigella, EHEC, and Yersinia. And showed a sensitivity of >90% for Campylobacter, Yersinia and EHEC, while Salmonella and Shigella obtained a dissentient sensitivity of <90%. No statistically significant difference could be detected for the Amplidiag® (p-value = 1.00) or for the BD-Max (p-value = 0.655) when compared to the available culture diagnostics. This study shows a strong argument to propose multiplex real-time PCR panels as a foundation for a reformed diagnostic method of intestinal pathogens. Based on a prominent improvement of factors such as; the time aspect and the performance of the method.

Place, publisher, year, edition, pages
Malmö universitet/Hälsa och samhälle , 2018. , p. 23
Keywords [sv]
culture diagnostics, intestinal pathogens, multiplex, real-time PCR
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:mau:diva-24711Local ID: 25643OAI: oai:DiVA.org:mau-24711DiVA, id: diva2:1486132
Educational program
HS Biomedicinsk laboratorievetenskap
Supervisors
Examiners
Available from: 2020-11-02 Created: 2020-11-02Bibliographically approved

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