Development and testing of new primers for LAMP detection of influenza B virus

Panhavoan Ky

Master's thesis

Abstract:

This study focuses on addressing challenges in the detection of the Influenza B virus (IBV), a significant threat causing flu-related morbidity in humans and infecting animals like seals. Current detection methods, such as Reverse Transcription Polymerase Chain Reaction (RT-PCR), are effective but require expensive equipment and time. The study proposes Loop-mediated Isothermal Amplification (LAMP) as a streamlined and rapid alternative for IBV detection, especially suitable for point-of-care settings. Primers were designed targeting a conserved region in the IBV Matrix 1 gene, ensuring sensitivity and specificity. Tests with both colorimetric and fluorescent products showed successful amplification within 12–19 minutes for fluorescent products and 40 minutes for colorimetric products, with visual changes indicating positive results. The assay was validated with negative, cross-reactivity, and positive controls, as well as asymptomatic volunteers, demonstrating its effectiveness and potential for enhancing IBV detection accessibility and efficiency.

Keywords

Influenza B virus, Primer construction, Loop-mediated Isothermal Amplification

Language used: English

Date on which the thesis was submitted / produced: 22. 4. 2024

Thesis defence

• Date of defence: 3. 6. 2024
• Supervisor: RNDr. Jiří Černý, Ph.D.
• Reader: Vojtěch Vacek, Mgr.

Czech University of Life Sciences Prague

Faculty of Tropical AgriSciences

Master programme / specializace:
Wildlife and Livestock Production, Management and Conservation / Sustainable Animal Production