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Utvärdering av Biosynex AMPLIQUICK® Fecal Parasitology multiplexpaneler för intestinal parasitdiagnostik med realtids-PCR: Jämförelse av förbehandlings- och extraktionsprotokoll samt metodologisk resultatjämförelse mot mikroskopi
Jönköping University, School of Health and Welfare, HHJ, Department of Clinical Diagnostics.
Jönköping University, School of Health and Welfare, HHJ, Department of Clinical Diagnostics.
2025 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Evaluation of the Biosynex AMPLIQUICK® Fecal Parasitology Multiplex Panels for the Detection of Protozoan and Helminth Infections with Real-Time PCR : A Comparative Analysis of Pretreatment and Extraction Protocols, and Methodological Differences Between PCR and Microscopy (English)
Abstract [sv]

Studien utvärderade Biosynex AMPLIQUICK® Fecal Parasitology multiplexpaneler för detektion av protozo- och helmintinfektioner med realtids-PCR. Utvärderingen utfördes genom en jämförelse av tre olika förbehandlingar samt två olika extraktioner på sparade frysta fecesprover, positivt påvisade för protozoer respektive helminter. Prospektiva patientprover användes till en metodologisk resultatjämförelse mellan realtids-PCR och mikroskopi. DNA extraherades automatiserat och analyserades med Biosynex AMPLIQUICK® PCR-strip. Databearbetning genomfördes i Excel.

Resultaten visade att Mikrobiologiska laboratoriets, Länssjukhuset Ryhov, enzymatiska samt fysikaliska rutinförbehandling i kombination med EZ1 DNA Bacteria Card v1.0 genererade lägst Cycle threshold-värden och högst detektionsfrekvens. Biosynex AMPLIQUICK® Fecal Pretreatment med mekanisk förbehandling uppvisade lägre prestanda. Mikroskopi och realtids-PCR visade god överensstämmelse. En högre andel positiva prover observerades för realtids-PCR, vilket antyder en högre känslighet för metoden.

Abstract [en]

This study evaluated the Biosynex AMPLIQUICK® Fecal Parasitology multiplex panels for the detection of protozoan and helminth infections using Real-Time PCR. The evaluation was conducted through a comparative analysis of three pretreatment protocols and two DNA extraction methods on positive preserved, fecal samples. The methodological comparison of results between Real-Time PCR and microscopy was performed with clinical samples. DNA extraction was performed using an automated system, and the analyses were conducted using the Biosynex AMPLIQUICK® PCR strip. Data processing was carried out with Microsoft Excel.

The results demonstrated that the enzymatic and physical routine pretreatment, used at the Department of Clinical Microbiology at Ryhov Hospital, in combination with the EZ1 DNA Bacteria Card v1.0 yielded the lowest Cycle threshold values and the highest detection rates. In contrast the Biosynex AMPLIQUICK® Fecal Pretreatment, which relied on mechanical lysis, showed lower performance. A high concordance was observed between microscopy and Real-Time PCR, although Real-Time PCR detected a higher proportion of positive samples, suggesting superior analytical sensitivity.

Place, publisher, year, edition, pages
2025. , p. 24
Keywords [en]
Intestinal Parasite Infection, Real-Time PCR, Microscopy, Pretreatment, Extraction
Keywords [sv]
Intestinala parasitinfektioner, realtids-PCR, mikroskopi, förbehandling, extraktion
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:hj:diva-68839OAI: oai:DiVA.org:hj-68839DiVA, id: diva2:1973759
Subject / course
HHJ, Biomedical Laboratory Science
Supervisors
Examiners
Available from: 2025-06-24 Created: 2025-06-19 Last updated: 2025-10-13Bibliographically approved

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