Development of a nucleic acid-based isothermal diagnostic test for border disease which causes losses in animal husbandry için kapak resmi
Development of a nucleic acid-based isothermal diagnostic test for border disease which causes losses in animal husbandry
Başlık:
Development of a nucleic acid-based isothermal diagnostic test for border disease which causes losses in animal husbandry
Yazar:
Ayaz Kök, Sanem, author.
Yazar Ek Girişi:
Fiziksel Tanımlama:
xi, 86 leaves: charts;+ 1 computer laser optical disc
Özet:
Border 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.
Tek Biçim Eser Adı:
Thesis (Master)--İzmir Institute of Technology: Biotechnology.

İzmir Institute of Technology: Biotechnology--Thesis (Master).
Elektronik Erişim:
Access to Electronic Versiyon.
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