Title:Bio-Membrane SELEX as a New Approach for Selecting ss-DNA
Aptamers that Bind to the Hydatid Cyst Laminated Layer
Volume: 23
Issue: 6
Author(s): Azar Balouti Dehkordi*, Hossein Khanahmad, Yadollah Azizi, Mina Mirian, Nader Pestehchian and Hossein Yousofi Darani
Affiliation:
- Department of Medical Parasitology and Mycology, Faculty of Medicine, Isfahan University of Medical
Sciences, Isfahan, Iran
Keywords:
Aptamer, SELEX, Echinococcus granulosus, Parasites, Helminthes, Echinococcosis.
Abstract:
Background: Hydatid cyst (HC) is the larval stage of the canine intestinal
tapeworm (cestode), Echinococcus granulosus. In addition to the high global economic
cost of livestock farming, the infection can lead to dangerous problems for human
health. Therefore, research into new diagnosis and treatment approaches is valuable.
This study is set out to explore aptamers that bind to HC antigens.
Methods: The similarity between HC genotype in sheep and humans was that sheep
HCs were collected and used as a biological membrane for aptamer selection. Four Bio-
Membrane SELEX rounds were conducted, and ssDNA aptamers were selected.
Selected aptamers' affinity and specificity to the laminated layer antigens were evaluated
using membrane staining by fluorescein primer as a probe. Biotinylated primer was used
as a probe for aptahistochemistry and dot blot techniques. Subsequently, cloning and
plasmid extraction was conducted. The affinity and specificity of sequenced aptamers
were examined with the dot blot method.
Results: Selected aptamers reacted with HC wall in aptahistochemistry,
aptahistofluorescent, and dot blot experiments. Following cloning and sequencing, 20
sequences were achieved. A strong reaction between HC total antigens and sequenced
aptamers has emerged in the dot blot method.
Conclusion: We propose a novel method to determine specific aptamers in this
investigation. Bio-Membrane SELEX could be assumed as a practical and sensitive
method for aptamer selection. Selected aptamers in this study possibly may be used for
specific HC antigens detection.
