DNA ligase photocrosslinked cryogenic column based biotinylation kit for viral hybridization and detection

Abstract

This study describes the development of cryogel columns prepared by ATP having ruthenium hapten cross-linker based photosensitive approach "ANADOLUCA" for the biotinylation of Crimean Congo Hemorrhagic Fever (CCHF) primers through 5' end as a first. In this process, which does not require ATP ligand, CCHF recognition has been performed using stable and quantum dots (QDs) based avidity nanoparticles cross-linked with streptavidin. p(HEMA-co-DNA ligase) photosensitive copolymerized cryogel column as a biotinylated continuous system have prepared using ATP ligand having photosensitive ruthenium chelate based hapten cross-linker and characterized. Then, the biotinylated efficiency of the prepared column has performed through continuous biotin binding without any need of ATP ligand having to Crimean congo hemorrhagic fever primer (CCHF). The efficiency of the biotinylated CCHF primer has investigated by polymerase chain reaction (PCR). The characteristic of a sustainable biotinylating column synthesized from the same ligand that we normally develop using DNA ligase while working with ATP in the DNA ligase biotinylation process is that it can biotinize without ATP. In addition, virus-identifying ssDNA was observed as fluorescence after avidity interaction with the QD combination for viral identification after biotinylation. Then, the fluorescence intensity changes through the hybridization of 3'-ssDNA complements have determined.