Aptamēru atlase pret plasmodium vivax laktātdehidrogenāzi un specifisko sekvenču validācija
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Latvijas Universitāte
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lav
Abstract
Malārija, ko izraisa Plasmodium ģints parazīti, ir smaga, dzīvībai bīstama infekcijas slimība. Pasaules Veselības organizācija (PVO) cenšas izskaust malāriju, veicot plašu skrīningu, tomēr pašreizējie diagnostikas rīki ir pārāk sarežģīti vai tiem trūkst jutīguma, lai efektīvi veiktu malārijas diagnostiku. Lai ierobežotu malārijas saslimtību ir nepieciešami rentabli, stabili un augstas jutības diagnostikas rīki. Aptamēri ir īsi DNS vai RNS oligonukleotīdi, kas telpiski saistās ar specifiskiem mērķiem, piemēram, dažādām olbaltumvielām un piedāvā vairākas priekšrocības salīdzinājumā ar antivielām, piemēram, mazāku izmēru, zemākas ražošanas izmaksas, ķīmisko stabilitāti un ilgāku glabāšanas laiku, padarot tos ideāli piemērotus aprūpes punktos (point-of-care) lietojamu biosensoru izstrādei malārijas noteikšanai. Šī darba ietvaros tika veikta aptamēru atlase, izmantojot proteīnu SELEX metodi kā rezultātā tika atlasīti seši aptamēri, kuru specifiskums pret mērķi tika pārbaudīts ar ELONA metodi. Kā arī tika testēta šo aptamēru stabilitāte cilvēka serumā.
Malaria, caused by Plasmodium parasites, is a severe, life-threatening infectious disease. The World Health Organization (WHO) aims to eliminate malaria through widespread screening; however, current diagnostic tools are either too complex or lack the sensitivity to effectively diagnose malaria. To control malaria incidence, cost-effective, stable, and highly sensitive diagnostic tools are needed. Aptamers are short, three-dimensional DNA or RNA single-strand oligonucleotides that bind to specific targets, such as various proteins, and offer several advantages over antibodies, such as smaller size, lower production costs, chemical stability, and longer shelf-life, making them ideal for developing point-of-care biosensors for malaria detection. In this work, aptamer selection was performed using the protein-SELEX method, resulting in the selection of six aptamers, whose specificity against the target was tested using the ELONA method. Additionally, the stability of these aptamers in human serum was tested.
Malaria, caused by Plasmodium parasites, is a severe, life-threatening infectious disease. The World Health Organization (WHO) aims to eliminate malaria through widespread screening; however, current diagnostic tools are either too complex or lack the sensitivity to effectively diagnose malaria. To control malaria incidence, cost-effective, stable, and highly sensitive diagnostic tools are needed. Aptamers are short, three-dimensional DNA or RNA single-strand oligonucleotides that bind to specific targets, such as various proteins, and offer several advantages over antibodies, such as smaller size, lower production costs, chemical stability, and longer shelf-life, making them ideal for developing point-of-care biosensors for malaria detection. In this work, aptamer selection was performed using the protein-SELEX method, resulting in the selection of six aptamers, whose specificity against the target was tested using the ELONA method. Additionally, the stability of these aptamers in human serum was tested.