Generic placeholder image

Current Molecular Medicine


ISSN (Print): 1566-5240
ISSN (Online): 1875-5666

Research Article

Bio-Membrane SELEX as a New Approach for Selecting ss-DNA Aptamers that Bind to the Hydatid Cyst Laminated Layer

Author(s): Azar Balouti Dehkordi*, Hossein Khanahmad, Yadollah Azizi, Mina Mirian, Nader Pestehchian and Hossein Yousofi Darani

Volume 23, Issue 6, 2023

Published on: 26 August, 2022

Page: [578 - 587] Pages: 10

DOI: 10.2174/1566524022666220516140451

Price: $65


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.

Keywords: Aptamer, SELEX, Echinococcus granulosus, Parasites, Helminthes, Echinococcosis.

« Previous
Garcia LS. Diagnostic medical parasitology. American Society for Microbiology Press 2006.
John DT, Petri WA Jr. Markell and Voge’s Medical Parasitology. Elsevier Health Sciences 2006.
Gollackner B, Längle F, Auer H, et al. Radical surgical therapy of abdominal cystic hydatid disease: Factors of recurrence. World J Surg 2000; 24(6): 717-21.
[] [PMID: 10773125]
Doğan R, Yüksel M, Cetin G, et al. Surgical treatment of hydatid cysts of the lung: Report on 1055 patients. Thorax 1989; 44(3): 192-9.
[] [PMID: 2705149]
Jenkins DJ, Romig T, Thompson RC. Emergence/re-emergence of Echinococcus spp.--a global update. Int J Parasitol 2005; 35(11-12): 1205-19.
[] [PMID: 16157340]
Parija SC. Textbook of medical parasitology, Protozoology & Helminthology. SciELO Brasil 2008.
Song KM, Lee S, Ban C. Aptamers and their biological applications. Sensors 2012; 12(1): 612-31.
[] [PMID: 22368488]
Lee JF, Hesselberth JR, Meyers LA, Ellington AD. Aptamer database. Nucleic Acids Res 2004; 32 (suppl_1): D95-D100.
[] [PMID: 14681367]
Ospina JD. Aptamers as a novel diagnostic and therapeutic tool and their potential use in parasitology. Biomedicine 2020; 40: 148-66.
Guerra-Pérez N, Ramos E, García-Hernández M, et al. Molecular and functional characterization of ssDNA aptamers that specifically bind Leishmania infantum PABP. PLoS One 2015; 10(10): e0140048.
[] [PMID: 26457419]
Barfod A, Persson T, Lindh J. In vitro selection of RNA aptamers against a conserved region of the Plasmodium falciparum erythrocyte membrane protein 1. Parasitol Res 2009; 105(6): 1557-66.
[] [PMID: 19693540]
Cheung YW, Dirkzwager RM, Wong WC, Cardoso J, D’Arc Neves Costa J, Tanner JA. Aptamer-mediated plasmodium-specific diagnosis of malaria. Biochimie 2018; 145: 131-6.
[] [PMID: 29080831]
Cheung YW, Kwok J, Law AW, Watt RM, Kotaka M, Tanner JA. Structural basis for discriminatory recognition of plasmodium lactate dehydrogenase by a DNA aptamer. Proc Natl Acad Sci 2013; 110(40): 15967-72.
[] [PMID: 24043813]
Iqbal A, Labib M, Muharemagic D, Sattar S, Dixon BR, Berezovski MV. Detection of cryptosporidium Parvum oocysts on fresh produce using DNA aptamers. PLoS One 2015; 10(9): e0137455.
[] [PMID: 26334529]
Ospina-Villa JD, Dufour A, Weber C, et al. Targeting the polyadenylation factor EhCFIm25 with RNA aptamers controls survival in Entamoeba histolytica. Sci Rep 2018; 8(1): 5720.
[] [PMID: 29632392]
Göringer HU, Homann M, Zacharias M, Adler A. RNA aptamers as potential pharmaceuticals against infections with African trypanosomes. Handb Exp Pharmacol 2006; 173: 375-93.
[] [PMID: 16594626]
Hall M III, Misra S, Chaudhuri M, Chaudhuri G. Peptide aptamer mimicking RAD51-binding domain of BRCA2 inhibits DNA damage repair and survival in Trypanosoma brucei. Microb Pathog 2011; 50(5): 252-62.
[] [PMID: 21296653]
Long Y, Qin Z, Duan M, et al. Screening and identification of DNA aptamers toward Schistosoma japonicum eggs via SELEX. Sci Rep 2016; 6(1): 24986.
[] [PMID: 27121794]
Ospina-Villa JD, López-Camarillo C, Castañón-Sánchez CA, Soto-Sánchez J, Ramírez-Moreno E, Marchat LA. Advances on aptamers against protozoan parasites. Genes 2018; 9(12): 584.
[] [PMID: 30487456]
Sefah K, Shangguan D, Xiong X, O’Donoghue MB, Tan W. Development of DNA aptamers using Cell-SELEX. Nat Protoc 2010; 5(6): 1169-85.
[] [PMID: 20539292]
Hammond JB, Kruger NJ. The bradford method for protein quantitation New Protein Techniques. Springer 1988; pp. 25-32.
Khademvatan S, Majidiani H, Foroutan M, Tappeh KH, Aryamand S, Khalkhali HR. Echinococcus granulosus genotypes in Iran: A systematic review. J Helminthol 2019; 93(2): 131-8.
[] [PMID: 29606162]
Lantero E, Belavilas-Trovas A, Biosca A, et al. Development of DNA aptamers against Plasmodium falciparum blood stages using cell-systematic evolution of ligands by exponential enrichment. J Biomed Nanotechnol 2020; 16(3): 315-34.
[] [PMID: 32493542]
Ellington AD, Szostak JW. In vitro selection of RNA molecules that bind specific ligands. Nature 1990; 346(6287): 818.
Tuerk C, Gold L. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 1990; 249(4968): 505-10.
Kouhpayeh S, Hejazi Z, Khanahmad H, Rezaei A. Real-time PCR: An appropriate approach to confirm ssDNA generation from PCR product in SELEX process. Iran J Biotechnol 2017; 15(2): 143-8.
[] [PMID: 29845062]
Bauer M, Macdonald J, Henri J, Duan W, Shigdar S. The application of aptamers for immunohistochemistry. Nucleic Acid Ther 2016; 26(3): 120-6.
[] [PMID: 26862683]
Zeng Z, Zhang P, Zhao N, et al. Using oligonucleotide aptamer probes for immunostaining of formalin-fixed and paraffin-embedded tissues. Mod Pathol 2010; 23(12): 1553-8.
[] [PMID: 20693984]
Chen Z, Wu Q, Chen J, Ni X, Dai J. A DNA aptamer based method for detection of SARS-CoV-2 nucleocapsid protein. Virol Sin 2020; 35(3): 351-4.
[] [PMID: 32451881]
Yang KA, Pei R, Stojanovic MN. In vitro selection and amplification protocols for isolation of aptameric sensors for small molecules. Methods 2016; 106: 58-65.
[] [PMID: 27155227]
Smirnov I, Shafer RH. Effect of loop sequence and size on DNA aptamer stability. Biochemistry 2000; 39(6): 1462-8.
[] [PMID: 10684628]
Cai S, Yan J, Xiong H, Liu Y, Peng D, Liu Z. Investigations on the interface of nucleic acid aptamers and binding targets. Analyst 2018; 143(22): 5317-38.
[] [PMID: 30357118]
Setlem K, Mondal B, Ramlal S, et al. Immuno affinity SELEX for simple, rapid, and cost-effective aptamer enrichment and identification against aflatoxin B1. Front Microbiol 2016; 7: 1909.
[PMID: 27990137]

Rights & Permissions Print Export Cite as
© 2023 Bentham Science Publishers | Privacy Policy