Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Development of an Advanced Lspr-Based Biosensor Chip for Rapid Detection of Border Disease Virus(01. Izmir Institute of Technology, 2023) Alakbarov, Abdullah; Bulmuş Zareie, Esma Volga; Tekin, Hüseyin CumhurThe Border Disease Virus (BDV) is responsible for causing fetal deathly infection, leading to annual occurrences of affected farms. BDV, along with other pestiviruses such as classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV), are known to cause major losses in stock farming. These losses can result in reproductive failure, expensive inspections, and other impacts on livestock health. The current detection methods of BDV include various techniques such as RT-PCR, ELISA, VNT, and immunofluorescence assays. These methods, although reliable, may require specialized equipment, time-consuming procedures, and laboratory facilities, making them less suitable for rapid on-site detection. Hence, it is imperative to employ diverse methodologies for detection of BDV. LSPR-based biosensors are a subset of plasmonic biosensors that exhibit numerous advantages for diverse applications. LSPR-based biosensors are particularly well-suited for the production of compact, practical devices for rapid, on-site detection of analytes. The aim of this study is to design and fabricate a biosensor chip utilizing LSPR technology for potential BDV detection. For this aim, glass surfaces were functionalized with gold nanorods modified with a BDV-specific primer sequence, complementary single-strand DNA sequence of 19 bases, and fabricated with PMMA microchannels. Different concentrations of target BDV-DNAsequence ranging from 0.01 pM to 100 nM were exposed to the channels, and the LSPR response was quantified using a Vis-NIR spectrometer. The limit of quantification of the biosensor chips was determined to be 10 pM, while the limit of detection was found to be less than or equal to 1 pM. The sensitivity of the biosensor chips was calculated to be 0.0567 nm/RIU. The dynamic range of the biochips lies between 10 pM to 100 pM.Master Thesis Development of a Nucleic Acid-Based Isothermal Diagnostic Test for Border Disease Which Causes Losses in Animal Husbandry(01. Izmir Institute of Technology, 2022) Ayaz Kök, Sanem; Taşkent Sezgin, Hümeyra; Meşe Özçivici, GülistanBorder disease is viral infection of ruminants, and it is associated with abortions, stillbirth, and birth of persistently infected (PI) lambs. It has a great potential to cause an outbreak and it is declared as one of the notifiable diseases of ruminants by World Organization for Animal Health (OIE). Border disease poses a threat against ruminant farming industry by causing major economic losses. Since there is no treatment or vaccine against border disease virus (BDV), early diagnosis and early isolation of infected animals is necessary. RT-qPCR is the gold-standard method for BDV identification, but it can only be applied by trained personnel in a laboratory with expensive instruments. There is a need for a point-of-care (POC) test, specifically designed for BDV. This thesis study aimed to develop a nucleic acid-based loop mediated isothermal amplification (LAMP) technique for BDV identification. LAMP is a nucleic acid identification technique that can be performed using 4-6 primers at a constant temperature with a Bst DNA polymerase. Firstly, multiple alignment of BDV sequences across the world was performed and most conserved region of genome was detected as 5’UTR. Then, three LAMP primer sets 1, 2a and 2b were designed to target 5’UTR. Designed primer sets were optimized in terms of temperature, fluorescent dye, primer mix, Mg2+ and enzyme concentration. After designation of optimum conditions, limit of detection (LOD) was determined for each primer set and their performances were compared. All primer sets have LOD equals to 2x104 copies/μl. Overall, primer set 1 and 2b has higher sensitivity and specificity compared to primer set 2a, therefore they are more suitable to be used for BDV identification with LAMP.