طراحي و بهينه سازي آپتامر جهت تشخيص ويروس هرپس سيمپلكس انساني نوع 1 و 2

Herpes simplex viruses 1 and 2 (HSV-1 and HSV-2), which cause oral and genital herpes, are preva‎lent worldwide, and epidemiological studies show an increase in infection rates in most countries. Type 1 (HSV-1) spreads primarily through oral contact and causes oral and genital infections. Type 2 (HSV-2) spreads through sexual contact and causes genital herpes. ELISA techniques, real-time PCR, and cytological tests are conventional methods for detection of these viruses, which are very expensive, time-consuming, and labor-intensive. Therefore, the main purpose of this study was to design a specific G-quadruplex aptamer for the simple and rapid detection of human herpes simplex virus types 1 and 2. In this study, a specific aptamer was designed using servers and bioinformatics software for the GD protein in HSV-1 and HSV-2 viruses. After eva‎luating the binding of the aptamer to the viral GD protein based on the stability scores of the secondary and tertiary structures and molecular docking, the aptamer (AptNR88) was selected and its binding to the target protein was confirmed experimentally using a colorimetric system with gold nanoparticles. In the laboratory section, gold nanoparticles with an average particle size of 30 nm were synthesized, and the aptamer AptNR88 was attached to gold nanoparticles through N atom bonds of adenine and guanine bases and electrostatic bonds. Then, the concentration of the selected aptamer was optimized for resistance against salt-induced aggregation in the colorimetric system, and colorimetric tests showed that the color change of gold nanoparticles from red to purple due to salt only occurs in the presence of the AptNR88-aptamer-virus complex after 15 minutes. These results confirmed the specific binding of AptNR88 to the GD of HSV-1 and HSV-2 viruses. It can be concluded that the designed aptamer for the detection of these two viruses is sufficiently sensitive and specific.

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